Method for treating a semiconductor device
49 claims: 49 independent, 0 dependent
- 1センサアレイを処理する方法 であって、 該センサアレイが複数のセンサを含み、該複数のセンサのうちの1つのセンサが、該センサアレイの表面に曝露されているセンサパッドを有し 、該センサがISFETセンサであり、該方法が、 少なくとも前記センサパッドを酸及び有機溶媒を含む洗浄溶液に曝露する 工程であって、該酸がスルホン酸を含む、工程 と、 該センサパッドから該洗浄溶液をすすぎ落とす 工程と、 を含む方法 。
- 2前記スルホン酸が、アルキルスルホン酸、アルキルアリールスルホン酸、またはそれらの組み合わせを含む、請求項 1 に記載の方法。
- 3前記アルキルアリールスルホン酸が、1~20個の炭素を有するアルキル基を含む、請求項 2 に記載の方法。
- 4前記アルキル基が、9~18個の炭素を有する、請求項 3 に記載の方法。
- 5前記アルキル基が、10~14個の炭素を有する、請求項 4 に記載の方法。
- 6前記アルキル基が、1~6個の炭素を有する、請求項 3 に記載の方法。
- 7センサアレイを処理する方法 であって、該 センサアレイが複数のセンサを含み、該複数のセンサのうちの1つのセンサがセンサパッドを含み、 該センサがISFETセンサであり、 ウェル構造が該センサアレイに対応するウェルアレイを画定し、該ウェルアレイのウェルが該センサパッドを曝露し、キャップが該センサアレイ及び該ウェル構造にわたって取り付けられかつ流体ポートを含み、空間が該キャップと該ウェル構造との間に画定され 、該方法が、 酸及び有機溶媒を含む洗浄溶液を、前記流体ポートを通して前記空間内に適用し、30秒~30分の第1の期間待機する 工程であって、該酸がスルホン酸を含む、工程 と、 塩基性溶液を、前記流体ポートを通して前記空間内に適用し、20秒~15分の第2の期間待機する 工程 と、 すすぎ溶液を、前記流体ポートを通して適用する 工程と、 を含む方法 。
- 8前記洗浄溶液を適用する 工程 、前記塩基性溶液を適用する 工程 、及び前記すすぎ溶液を適用する 工程 を繰り返すことを更に含む、請求項 7 に記載の方法。
- 9センサアレイを処理する方法 であって、該 センサアレイが複数のセンサを含み、該複数のセンサのうちの1つのセンサがセンサパッドを含み、 該センサがISFETセンサであり、 ウェル構造が該センサアレイに対応するウェルアレイを画定し、該ウェルアレイのウェルが該センサパッドを曝露して おり、該方法が、 酸及び有機溶媒を含む洗浄溶液を少なくとも前記センサパッドに適用し、30秒~30分の第1の期間待機する 工程であって、該酸がスルホン酸を含む、工程 と、 少なくとも前記センサパッドを低沸点有機溶媒ですすぐ 工程 と、 前記センサアレイを乾燥させる 工程と、 を含む方法 。
- 10前記低沸点有機溶媒ですすいだ後であり、かつ乾燥させる前に、水ですすぐ 工程 を更に含む、請求項 9 に記載の方法。
- 11キャップを前記センサアレイ及び前記ウェルアレイにわたって取り付ける 工程 を更に含み、該キャップが流体ポートを含み、空間が、前記流体ポートと流体連通している状態で該キャップと該ウェルアレイとの間に画定されている、請求項 9 に記載の方法。
- 12前記スルホン酸が、メタンスルホン酸、エタンスルホン酸、プロパンスルホン酸、ブタンスルホン酸、またはそれらの組み合わせを含む、請求項 1~11 のいずれか一項に記載の方法。
- 13前記スルホン酸が、ドデシルベンゼンスルホン酸を含む、請求項 1~11 のいずれか一項に記載の方法。
- 14前記スルホン酸が、パラトルエンスルホン酸を含む、請求項 1~11 のいずれか一項に記載の方法。
- 15前記洗浄溶液が、10mM~500mMの前記酸を含む、請求項 1~14 のいずれか一項に記載の方法。
- 16前記洗浄溶液が、50mM~250mMの前記酸を含む、請求項 15 に記載の方法。
- 17前記洗浄溶液が、0.5重量%~25重量%の前記酸を含む、請求項 1~14 のいずれか一項に記載の方法。
- 18前記洗浄溶液が、1重量%~10重量%の前記酸を含む、請求項 17 に記載の方法。
- 19前記洗浄溶液が、2.5重量%~5重量%の前記酸を含む、請求項 18 に記載の方法。
- 20前記有機溶媒が、非極性である、請求項 1~19 のいずれか一項に記載の方法。
- 21前記有機溶媒が、36°C~345°Cの範囲の標準沸点を有する、請求項 1~20 のいずれか一項に記載の方法。
- 22前記標準沸点が、65°C~275°Cの範囲にある、請求項 21 に記載の方法。
- 23前記標準沸点が、65°C~150°Cの範囲にある、請求項 22 に記載の方法。
- 24前記標準沸点が、150°C~220°Cの範囲にある、請求項 22 に記載の方法。
- 25前記有機溶媒が、6~24個の炭素を有するアルカンである、請求項 1~24 のいずれか一項に記載の方法。
- 26前記アルカンが、6~14個の炭素を有する、請求項 25 に記載の方法。
- 27前記アルカンが、6~9個の炭素を有する、請求項 26 に記載の方法。
- 28前記アルカンが、10~14個の炭素を有する、請求項 26 に記載の方法。
- 29前記有機溶媒が、極性非プロトン性溶媒である、請求項 1~19 のいずれか一項に記載の方法。
- 30前記極性非プロトン性溶媒が、テトラヒドロフラン、酢酸エチル、アセトン、ジメチルホルムアミド、アセトニトリル、ジメチルスルホキシド、N-メチルピロリドン、またはそれらの組み合わせを含む、請求項 29 に記載の方法。
- 31前記センサパッドを前記洗浄溶液に曝露しながら、前記センサパッド及び前記洗浄溶液を加熱する 工程 を更に含む、請求項 1~30 のいずれか一項に記載の方法。
- 32加熱する 工程 が、35°C~70°Cの範囲の温度で加熱することを含む、請求項 31 に記載の方法。
- 33前記温度が、40°C~55°Cの範囲にある、請求項 32 に記載の方法。
- 34曝露する 工程 が、30秒~30分の範囲の期間曝露することを含む、請求項 1~33 のいずれか一項に記載の方法。
- 35前記期間が、30秒~10分の範囲にある、請求項 34 に記載の方法。
- 36前記期間が、30秒~5分の範囲にある、請求項 35 に記載の方法。
- 37前記期間が、1分~3分の範囲にある、請求項 36 に記載の方法。
- 38少なくとも前記センサパッドを塩基性溶液に曝露する 工程 を更に含む、 1~37 のいずれか一項に記載の方法。
- 39前記塩基性溶液が、0.005M~1.5Mの水酸化ナトリウムを含む、請求項 38 に記載の方法。
- 40前記塩基性溶液が、0.01M~1.0Mの水酸化ナトリウムを含む、請求項 39 に記載の方法。
- 41前記塩基性溶液が、0.01M~0.5Mの水酸化ナトリウムを含む、請求項 40 に記載の方法。
- 42少なくとも前記センサパッドを前記塩基性溶液に曝露する 工程 が、前記センサパッドを前記洗浄溶液に曝露した後に起こる、請求項 38 に記載の方法。
- 43少なくとも前記センサパッドを前記洗浄溶液に曝露する 工程 及び少なくとも前記センサパッドを前記塩基性溶液に曝露する 工程 を繰り返すことを更に含む、請求項 38 に記載の方法。
- 44すすぎ落とす 工程 が、水及び前記有機溶媒と混和可能なすすぎ有機溶媒ですすぐことを含む、請求項 1~43 のいずれか一項に記載の方法。
- 45すすぎ落とす 工程 が、25°C~100°Cの範囲の標準沸点を有する低沸点有機溶媒ですすぐことを含む、請求項 1~44 のいずれか一項に記載の方法。
- 46前記低沸点有機溶媒が、50°C~100°Cの範囲の標準沸点を有する、請求項 45 に記載の方法。
- 47前記低沸点有機溶媒が、アルコールを含む、請求項 45 に記載の方法。
- 48前記アルコールが、エタノールまたはイソプロパノールを含む、請求項 47 に記載の方法。
- 49すすぎ落とす 工程 が、水ですすぐことを含む、請求項 1~48 のいずれか一項に記載の方法。
Independent claims49
90 paragraphs, as filed
Cross-reference to related applications (s) This application claims the benefit of US Provisional Patent Application No. 61 / 806,603 filed March 29, 2013, which is hereby incorporated by reference in its entirety. Incorporated into the book.
This application claims the interests of US Provisional Patent Application No. 61 / 817,805 filed April 30, 2013, which is incorporated herein by reference in its entirety.
The present disclosure generally relates to methods for processing sensor arrays, sensor arrays formed by such methods, and solutions for use in such methods.
Arrays of sensors formed on semiconductor substrates are increasingly being used in fields such as analytical chemistry and molecular biology. For example, if the analyte is captured on or near the sensor pad of the sensor array, the reactants or by-products of the reaction associated with the analyte will be detected and used to provide information about the analyte. Can be elucidated. Specifically, such sensor arrays have been found to be used in gene analysis such as gene sequence or quantitative amplification.
During manufacturing, various semiconductor processing techniques can change the properties of the surface of the sensor array and the surface of the well structure around the sensor array. Such processing may also leave residues on the surface. Altered surface chemistry and residues can prevent or limit capture of the analyte in close proximity to the sensor. Therefore, the effectiveness of such a sensor array is reduced, and the signal obtained from such a sensor array may or may not contain erroneous data.
In one aspect, the sensor device comprising the sensor array and optionally the well array corresponding to the sensor array, or the cap attached to the sensor array, can be treated with a wash solution. The cleaning solution can contain an organic solvent and an acid such as sulfonic acid, for example alkylbenzene sulfonic acid. The sensor device can be further treated with a basic solution such as NaOH solution or rinsed with a low boiling organic solvent or water. Optionally, the sensor device can be dried.<u style="single">The basic features and various aspects of the present invention are listed below.</u><u style="single">[1]</u><u style="single"> A method of processing the sensor array, including: the sensor array comprises a plurality of sensors, one of the sensors having a sensor pad exposed on the surface of the sensor array:</u><u style="single"> At least exposing the sensor pad to a cleaning solution containing an acid and an organic solvent</u><u style="single"> Rinse the cleaning solution from the sensor pad.</u><u style="single">[2]</u><u style="single"> The method according to [1], wherein the acid comprises a sulfonic acid.</u><u style="single">[3]</u><u style="single"> The method according to [2], wherein the sulfonic acid comprises an alkylsulfonic acid, an alkylarylsulfonic acid, or a combination thereof.</u><u style="single">[4]</u><u style="single"> The method according to [3], wherein the alkylaryl sulfonic acid contains an alkyl group having 1 to 20 carbons.</u><u style="single">[5]</u><u style="single"> The method according to [4], wherein the alkyl group has 9 to 18 carbons.</u><u style="single">[6]</u><u style="single"> The method according to [5], wherein the alkyl group has 10 to 14 carbons.</u><u style="single">[7]</u><u style="single"> The method according to [4], wherein the alkyl group has 1 to 6 carbons.</u><u style="single">[8]</u><u style="single"> The item according to any one of [2] to [7] and [45] to [49], wherein the sulfonic acid contains methanesulfonic acid, ethanesulfonic acid, propanesulfonic acid, butanesulfonic acid, or a combination thereof. the method of.</u><u style="single">[9]</u><u style="single"> The method according to any one of [2] to [7] and [45] to [49], wherein the sulfonic acid contains dodecylbenzenesulfonic acid.</u><u style="single">[10]</u><u style="single"> The method according to any one of [2] to [7] and [45] to [49], wherein the sulfonic acid contains paratoluenesulfonic acid.</u><u style="single">[11]</u><u style="single"> The method according to any one of [1] to [10] and [45] to [49], wherein the washing solution contains the acid of 10 mM to 500 mM.</u><u style="single">[12]</u><u style="single"> The method according to [11], wherein the cleaning solution comprises 50 mM to 250 mM of the acid.</u><u style="single">[13]</u><u style="single"> The method according to any one of [1] to [10] and [45] to [49], wherein the washing solution contains 0.5% by weight to 25% by weight of the acid.</u><u style="single">[14]</u><u style="single"> The method according to [13], wherein the washing solution contains 1% by weight to 10% by weight of the acid.</u><u style="single">[15]</u><u style="single"> The method according to [14], wherein the washing solution contains 2.5% by weight to 5% by weight of the acid.</u><u style="single">[16]</u><u style="single"> The method according to any one of [1] to [15] and [45] to [49], wherein the organic solvent is non-polar.</u><u style="single">[17]</u><u style="single"> The method according to any one of [1] to [16] and [45] to [49], wherein the organic solvent has a standard boiling point in the range of 36 ° C to 345 ° C.</u><u style="single">[18]</u><u style="single"> The method according to [17], wherein the standard boiling point is in the range of 65 ° C to 275 ° C.</u><u style="single">[19]</u><u style="single"> The method according to [18], wherein the standard boiling point is in the range of 65 ° C to 150 ° C.</u><u style="single">[20]</u><u style="single"> The method according to [18], wherein the standard boiling point is in the range of 150 ° C to 220 ° C.</u><u style="single">[21]</u><u style="single"> The method according to any one of [1] to [20] and [45] to [49], wherein the organic solvent is an alkane having 6 to 24 carbons.</u><u style="single">[22]</u><u style="single"> The method according to [21], wherein the alkane has 6 to 14 carbons.</u><u style="single">[23]</u><u style="single"> The method according to [22], wherein the alkane has 6 to 9 carbons.</u><u style="single">[24]</u><u style="single"> The method according to [22], wherein the alkane has 10 to 14 carbons.</u><u style="single">[25]</u><u style="single"> The method according to any one of [1] to [15] and [45] to [49], wherein the organic solvent is a polar aprotic solvent.</u><u style="single">[26]</u><u style="single"> The method according to [25], wherein the polar aprotic solvent comprises tetrahydrofuran, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide, N-methylpyrrolidone, or a combination thereof.</u><u style="single">[27]</u><u style="single"> The item according to any one of [1] to [26] and [45] to [49], further comprising heating the sensor pad and the cleaning solution while exposing the sensor pad to the cleaning solution. Method.</u><u style="single">[28]</u><u style="single"> The method according to [27], wherein heating comprises heating at a temperature in the range of 35 ° C to 70 ° C.</u><u style="single">[29]</u><u style="single"> The method according to [28], wherein the temperature is in the range of 40 ° C to 55 ° C.</u><u style="single">[30]</u><u style="single"> The method according to any one of [1] to [29] and [45] to [49], wherein the exposure comprises exposure for a period ranging from 30 seconds to 30 minutes.</u><u style="single">[31]</u><u style="single"> The method according to [30], wherein the period is in the range of 30 seconds to 10 minutes.</u><u style="single">[32]</u><u style="single"> The method according to [31], wherein the period is in the range of 30 seconds to 5 minutes.</u><u style="single">[33]</u><u style="single"> The method according to [32], wherein the period is in the range of 1 minute to 3 minutes.</u><u style="single">[34]</u><u style="single"> The method according to any one of [1] to [33] and [45] to [49], further comprising exposing at least the sensor pad to a basic solution.</u><u style="single">[35]</u><u style="single"> The method according to [34], wherein the basic solution comprises 0.005M to 1.5M sodium hydroxide.</u><u style="single">[36]</u><u style="single"> The method according to [35], wherein the basic solution contains 0.01 M to 1.0 M sodium hydroxide.</u><u style="single">[37]</u><u style="single"> The method according to [36], wherein the basic solution contains 0.01 M to 0.5 M sodium hydroxide.</u><u style="single">[38]</u><u style="single"> The method according to [34], wherein at least exposing the sensor pad to the basic solution occurs after exposing the sensor pad to the cleaning solution.</u><u style="single">[39]</u><u style="single"> The method according to [34], further comprising repeatedly exposing at least the sensor pad to the cleaning solution and at least exposing the sensor pad to the basic solution.</u><u style="single">[40]</u><u style="single"> The method according to any one of [1] to [39] and [45] to [49], wherein rinsing comprises rinsing with water and a rinse organic solvent miscible with the organic solvent.</u><u style="single">[41]</u><u style="single"> In any one of [1] to [40] and [46] to [50], rinsing involves rinsing with a low boiling organic solvent having a standard boiling point in the range of 25 ° C to 100 ° C. The method described.</u><u style="single">[42]</u><u style="single"> The method according to [41], wherein the low boiling organic solvent has a standard boiling point in the range of 50 ° C to 100 ° C.</u><u style="single">[43]</u><u style="single"> The method according to [41], wherein the low boiling organic solvent comprises alcohol.</u><u style="single">[44]</u><u style="single"> The method according to [43], wherein the alcohol comprises ethanol or isopropanol.</u><u style="single">[45]</u><u style="single"> The method according to any one of [1] to [44] and [46] to [50], wherein rinsing involves rinsing with water.</u><u style="single">[46]</u><u style="single"> The sensor array contains a plurality of sensors, one of the sensors includes a sensor pad, a well structure defines a well array corresponding to the sensor array, and the wells of the well array provide the sensor pad. A method of processing the sensor array, including the following, where the cap is attached across the sensor array and the well structure and includes a fluid port and a space is defined between the cap and the well structure:</u><u style="single"> A wash solution containing an acid and an organic solvent is applied into the space through the fluid port and waits for a first period of 30 seconds to 30 minutes.</u><u style="single"> The basic solution is applied into the space through the fluid port and waits for a second period of 20 seconds to 15 minutes.</u><u style="single"> Apply the rinse solution through the fluid port.</u><u style="single">[47]</u><u style="single"> The method according to [46], further comprising applying the washing solution, applying the basic solution, and applying the rinsing solution repeatedly.</u><u style="single">[48]</u><u style="single"> The sensor array contains a plurality of sensors, one of the sensors includes a sensor pad, a well structure defines a well array corresponding to the sensor array, and the wells of the well array provide the sensor pad. How to handle an exposed sensor array that includes:</u><u style="single"> Applying a cleaning solution containing an acid and an organic solvent to at least the sensor pad and waiting for a first period of 30 seconds to 30 minutes.</u><u style="single"> At least rinse the sensor pad with a low boiling organic solvent and</u><u style="single"> Drying the sensor array.</u><u style="single">[49]</u><u style="single"> The method according to [48], further comprising rinsing with water after rinsing with the low boiling organic solvent and before drying.</u><u style="single">[50]</u><u style="single"> It further comprises mounting the cap over the sensor array and the well array, the cap comprising a fluid port, and a space defined between the cap and the well array with fluid communication with the fluid port. The method described in [48].</u><u style="single">[51]</u><u style="single"> A sensor device that is processed by the method according to any one of [1] to [42].</u><u style="single">[52]</u><u style="single"> A solution containing 2.5% to 5% by weight dodecylbenzenesulfonic acid and a non-polar linear alkane having 6 to 18 carbons.</u><u style="single">[53]</u><u style="single"> With a nucleotide solution containing at least one nucleotide type,</u><u style="single"> With polymer particles</u><u style="single"> With a washing solution containing 0.5% to 20% by weight sulfonic acid and an organic solvent having a standard boiling point in the range of 65 ° C to 275 ° C.</u><u style="single">Including, kit.</u>
By referring to the accompanying drawings, the present disclosure can be better understood and many features and advantages thereof will be apparent to those skilled in the art.
<figref num="1">Includes a diagram of an exemplary measurement system.</figref><figref num="2">Includes diagrams of exemplary measurement components.</figref><figref num="3">Includes a diagram of an exemplary array of measurement components.</figref><figref num="4">Includes an exemplary well arrangement diagram.</figref><figref num="5">Includes illustrations of exemplary wells and sensor arrangements.</figref><figref num="6">Includes a diagram of an exemplary sensor device.</figref><figref num="7">Includes a diagram of an exemplary sensor device.</figref><figref num="8">Includes a flow chart showing an exemplary method.</figref><figref num="9">Includes a flow chart showing an exemplary method.</figref><figref num="10">Includes a flow chart showing an exemplary method.</figref><figref num="11">Includes a diagram of an exemplary method for preparing an sequencing device.</figref>
Use the same reference symbols in different drawings to indicate similar or identical items.
In one exemplary embodiment, the method of processing the sensor array comprises applying the cleaning solution to the sensor array and rinsing the cleaning solution from the sensor array. Specifically, the cleaning solution contains an organic solvent such as sulfonic acid and an acid. The sulfonic acid may include alkyl or alkylaryl sulfonic acid. In one example, the alkyl or alkylaryl sulfonic acid can have an alkyl group having 9-18 carbons. For example, the sulfonic acid may include sulfonated dodecylbenzene. The organic solvent can be a non-polar solvent or an aprotic polar solvent. In one example, the organic solvent may have a standard boiling point in the range of 65 ° C to 275 ° C. In one example, the organic solvent comprises heptane. In another example, the organic solvent comprises undecane. In a further embodiment, the organic solvent comprises dimethylformamide, acetonitrile, dimethyl sulfoxide, or a combination thereof. After treatment with the wash solution, the sensor array can be rinsed with the rinse solution. In one example, the rinse solution comprises a low boiling organic solvent such as, for example, ethanol or an alcohol which is isopropanol. In another example, the rinse solution may contain water. The method may further comprise applying a basic solution or a weak acid solution after the application of the wash solution and before rinsing. The basic solution may have a pH of at least 7 such as at least 8 and may contain a strong base such as sodium hydroxide. In one example, the washing solution and the basic solution may be applied repeatedly, such as 2 times or more, 3 times or more, or 4 times or more, but generally 10 times or less.
In certain embodiments, the wash solution is applied to the sensor pads of at least one sensor array. The sensor array may include multiple sensors. The sensors in the sensor array may include sensor pads. Optionally, the well structure can be defined across the sensor array and can include multiple wells corresponding to the sensor pads of the sensor array. The wells of the well array can expose the sensor pad of the sensor. Optionally, a cap containing at least one fluid port can be placed or mounted across the sensor array and well structure. The space for the fluid can be defined between the cap and the well structure or sensor array and can communicate with the fluid port of the cap. The wash solution can be applied through the fluid port into the space between the cap and the well structure. Optionally, the basic solution can be applied into the space between the cap and the well structure through the fluid port after application of the wash solution. The application of the wash solution after the basic solution may be repeated at least 2 times or 3 more times. The rinse solution can be applied, for example, through a fluid port containing alcohol or water. Optionally, the system can be dried.
In another exemplary embodiment, the sensor array comprises a plurality of sensors. One of the sensors includes a sensor pad. The well structure includes a well array that is arranged across the sensor array and corresponds operably to the sensor array. The wells of the well array expose the sensor pad of the sensor. A wash solution containing an acid such as sulfonic acid and an organic solvent is applied to the sensor array for a period of 30 seconds to 30 minutes. The sensor array may be rinsed with a rinse solution. In one example, the rinse solution may contain a low boiling organic solvent such as alcohol. The sensor array may be rinsed at least once with a low boiling organic solvent. In another embodiment or in addition, the sensor array may be rinsed with water such as deionized water. The sensor array can be dried and caps can be attached across the well structure and sensor array. The cap may include at least one fluid port. The space is defined between the cap and the sensor array or well structure and communicates with the fluid port.
In certain embodiments, the sensor system includes a flow cell in which the sensor array is installed, includes a communication circuit that is in electronic communication with the sensor array, and includes a container and fluid control that communicates fluid with the flow cell. In one embodiment, FIG. 1 shows an enlarged and cross-sectional view of the flow cell 100 and shows a portion of the flow chamber 106. The reagent stream 108 flows over the surface of the well array 102, where the reagent stream 108 flows over the open ends of the wells of the well array 102. The well array 102 and the sensor array 105 can together form an integration unit that forms the low wall (or floor) of the flow cell 100. The reference electrode 104 can be fluidly connected to the flow chamber 106. Further, the flow cell cover 130 seals the flow chamber 106 to accommodate the reagent flow flow 108 within a limited area.
FIG. 2 shows an enlarged view of the well 201 and the sensor 214, as shown in 110 of FIG. Well volume, shape, aspect ratio (bottom width to well depth ratio, etc.), and other dimensional features depend on the nature of the reaction that occurs and the reagents, by-products, or labeling techniques used (if any). Can be selected based on. The sensor 214 may be a chemical field effect transistor (ChemFET), more specifically an ion sensitive FET (ISFET) in which the floating gate 218 has a sensor plate 220 optionally separated from the well interior by a material layer 216. .. In addition, the conductive layer (not shown) can be placed on the sensor plate 220. In one embodiment, the material layer 216 includes an ion-sensitive material layer. The material layer 216 may be, for example, an oxide of zirconium, hafnium, tantalum, aluminum, or a ceramic layer such as titanium, especially a nitride of titanium. In one embodiment, the material layer 216 can have a thickness in the range of 5 nm to 100 nm, for example, the range of 10 nm to 70 nm, the range of 15 nm to 65 nm, or even more in the range of 20 nm to 50 nm. Together, the sensor plate 220 and the material layer 216 form a sensor pad.
The material layer 216 is shown as extending beyond the limits of the FET components shown, but the material layer 216 extends along the bottom of the well 201 and optionally along the wall of the well 201. can do. The sensor 214 can correspond to the amount of charge 224 present on the material layer 216 facing the sensor plate 220 (it can generate an output signal for the amount of charge 224). Changes in charge 224 can change the current between the power supply 221 and drain 222 of the chemFET. The chemFET can then be used directly to provide a current-based output signal, or indirectly with additional circuitry to provide a voltage-based output signal. Reactants, wash solutions, and other reagents can be moved into or out of the wells by the diffusion mechanism 240.
In one embodiment, the reaction performed within the well 201 can be an analytical reaction, identifying or determining the characteristics or properties of the analyte of interest. Such reactions can produce by-products directly or indirectly, which affect the amount of charge adjacent to the sensor plate 220. If such by-products are produced in small amounts, deteriorate rapidly, or react with other constituents, multiple replications of the same analyte will be produced at well 201 in order to increase the output signal produced. It can be analyzed at the same time. In one embodiment, multiple replicas of the analyte can be attached to the solid support 212 either before or after deposition in well 201. In one embodiment, the solid phase support 212 can be particles such as polymer particles or inorganic particles. In another embodiment, the solid phase support 212 may be a polymer matrix, such as a hydrophilic polymer matrix, such as a hydrogel matrix.
Well 201 can be defined by a wall structure, which wall structure can be formed from one or more layers of material. In one embodiment, the wall structure extends from the bottom surface to the top surface of the well in a range of 0.01 micrometers to 10 micrometers, eg, a range of 0.05 micrometers to 10 micrometers, a range of 0.1 micrometers to 10 micrometers, It can have a thickness in the range of 0.3 micrometers to 10 micrometers, or a range of 0.5 micrometers to 6 micrometers, and the like. Specifically, the thickness may range from 0.01 micrometer to 1 micrometer, for example, from 0.05 micrometer to 0.5 micrometer, or from 0.05 micrometer to 0.3 micrometer. Well 201 can have a characteristic diameter, which is defined as the square root of 4 times the cross-sectional area (A) divided by Pi (eg, sqrt (4 * A / π)), 5 It may be less than or equal to, for example, 3.5 micrometers or less, 2.0 micrometers or less, 1.6 micrometers or less, 1.0 micrometer or less, 0.8 micrometer or less, or even 0.6 micrometer or less. Well 201 can have a characteristic diameter of at least 0.01 micrometer.
Although FIG. 2 shows a single-layer wall structure and a single-layer material layer 216, the system can include one or more wall structural layers, one or more conductive layers, or one or more material layers. For example, the wall structure can be formed from a layer containing an oxide of silicon or TEOS, or one or more layers containing a nitride of silicon.
In the particular embodiment shown in FIG. 3, the system 300 includes a well wall structure 302, which array of wells 304 is located on or operably linked to the sensor pad of the sensor array. To define. The well wall structure 302 defines the top surface 306. The bottom surface 308 associated with the well is located on the sensor pad of the sensor array. The well wall structure 302 defines a side wall 310 between the top surface 306 and the bottom surface 308. As mentioned above, the material layer in contact with the sensor pads of the sensor array extends along the underside 308 of the wells of the array of wells 304, or along at least part of the wall 310 defined by the well wall structure 302. can do. The material layer does not have to be present on the upper surface 306. Specifically, the polymer matrix can be placed in the wells of the array of wells 304. The top surface 306 may be substantially free of polymer matrix. For example, the top surface 306 can include regions where the polymer matrix is absent, such as at least 70% of the total region, at least 80% of the total region, at least 90% of the total region, or approximately 100% of the total region.
Although the wall surface of FIG. 2 is shown to extend substantially vertically and outward, the wall surface can extend in different directions and have different shapes. Substantially perpendicular means extending in the direction having components perpendicular to the surface defined by the sensor pad. For example, as shown in FIG. 4, the well wall 402 extends vertically and can be parallel to the vertical component 412 of the surface defined by the sensor pad. In another embodiment, the wall surface 404 can extend substantially vertically in the outward direction away from the sensor pad to provide the well with an opening larger than the area of the underside of the well. As shown in FIG. 4, the wall surface 404 extends in a direction having a vertical component parallel to the vertical component 412 of the surface 414. In an alternative embodiment, the wall surface 406 extends substantially perpendicular to the medial direction to provide a smaller opening area than the area underneath the well. The wall surface 406 extends in a direction having components parallel to the vertical component 412 of the surface 414.
Although surfaces 402, 404, or 406 are represented by straight lines, some semiconductor or CMOS manufacturing processes can result in structures with non-linear shapes. Specifically, the wall surface, such as the wall surface 408, and the upper surface, such as the upper surface 410, may have an arched shape or may have various non-linear shapes. The structures and devices shown herein are shown as having linear layers, surfaces, or shapes, but the actual layers, surfaces, or shapes resulting from the semiconductor process may differ to some extent, if any. Some include non-linear and arched variants of the illustrated embodiments.
FIG. 5 includes a diagram of an exemplary well with an ion-sensitive material layer. For example, the well structure 502 can define an array of wells, eg, exemplary wells 504, 506, or 508. The wells (504, 506, or 508) can be operably connected to, or connected to, an underlying sensor (not shown). An exemplary well 504 comprises an ion-sensitive material layer 510, which defines the bottom of the well 504 and extends within the structure 502. Although not shown in FIG. 5, a conductive layer, such as a gate, for example, a floating gate of an ion-sensitive field effect transistor, may be present under the ion-sensitive material layer 510.
In another embodiment, as indicated by well 506, the ion-sensitive material layer 512 can define the bottom of well 506 without extending within structure 502. In a further embodiment, the well 508 can include an ion-sensitive layer 514, which extends along at least a portion of the side wall 516 of the well 508 defined by the structure 502. As mentioned above, the ion-sensitive material layer 512 or 514 may be present on the conductive layer or on the gate of the underlying electronic device.
As shown in FIG. 6, the sensor device can include a sensor array 602 formed in the semiconductor substrate. The well structure is defined across the sensor array. The cap 604 is attached to a well structure or sensor array 602. For example, the cap 604 can be glued to the sensor array or well structure 602 using an adhesive. Cap 604 includes fluid ports 606 or 608, which communicate fluid with the flow cell 610 defined between the cap 604 and the sensor array and well array 602. The fluid applied to one of the fluid ports 606 or 608 can flow through the flow cell 610 and optionally out of the outer port 608 or 606. In another embodiment shown in FIG. 7, the cap 702 can include fluid ports 704 and 706 to define a larger flow space. The cap can be glued over a well array or sensor array, for example using an adhesive.
As shown in Method 800 of FIG. 8, the sensor array can be exposed to a wash solution containing an acid, a sulfonic acid, a phosphonic acid, or a combination thereof, as shown in 802. The cleaning solution can further contain an organic solvent.
Illustrative sulfonic acids include alkyl sulfonic acids, alkylaryl sulfonic acids, or combinations thereof. Exemplary alkyl sulfonic acids include alkyl groups having 1 to 18 carbons, 1 to 14 carbons, 1 to 10 carbons, 1 to 5 carbons, and the like. In another example, the alkyl group of the alkyl sulfonic acid has 10-14 carbons. For example, alkyl sulfonic acid can include methane sulfonic acid, ethane sulfonic acid, propane sulfonic acid, butane sulfonic acid, or a combination thereof. In another embodiment, the alkyl group can be functionalized, for example, with a terminal functional group opposite the sulfonic acid functional group. Examples of the functionalized alkyl sulfonic acid include an alkyl sulfonic acid functionalized with a terminal amino group, taurine and the like. In a further embodiment, the alkyl group of the sulfonic acid can be halogenated, fluorinated, etc.
In a further embodiment, the sulfonic acid comprises an alkylaryl sulfonic acid. Alkylaryl sulfonic acids, such as alkylbenzene sulfonic acids, can have an alkyl group having 1 to 20 carbons. For example, an alkyl group can have 9-18 carbons, 10-14 carbons, and the like. In one particular embodiment, the alkylaryl sulfonic acid comprises dodecylbenzene sulfonic acid. The dodecylbenzene sulfonic acid can be a purified form of a dodecylbenzene sulfonic acid having an alkylaryl sulfonic acid having an alkyl group having 12 carbons, such as at least 90%, at least 95% and the like. Alternatively, the dodecylbenzene sulfonic acid can include a blend of alkylbenzene sulfonic acids having an alkyl group having an average of 12 carbons. Alkylaryl sulfonic acids can be alkylated with a blend of positions along the alkyl chain. In another example, the alkyl group can have 1-6 carbons. For example, alkylaryl sulfonic acid may include toluene sulfonic acid.
The cleaning solution may have an acid, sulfonic acid, etc. at a concentration of 10 mM to 500 mM. For example, the wash solution can have an acid concentration of 50 mM to 250 mM. In another example, the wash solution comprises 0.5% to 25% by weight of acid, sulfonic acid, etc. For example, the washing solution may contain 1% to 10% by weight of acid, sulfonic acid and the like, 2.5% by weight to 5% by weight of acid, sulfonic acid and the like.
The organic solvent in the washing solution is a non-aqueous solvent that provides solubility in an acid (eg, sulfonic acid) to at least the above concentrations. In one example, the organic solvent can be aprotic. The organic solvent can be a non-polar organic solvent. In another example, the organic solvent can be a polar aprotic solvent. In one example, the organic solvent may have a standard boiling point in the range of 36 ° C to 345 ° C. For example, the standard boiling point can be in the range of 65 ° C to 275 ° C. In another example, the standard boiling point can be in the range of 65 ° C to 150 ° C. Alternatively, the standard boiling point is in the range of 150 ° C to 220 ° C.
In certain embodiments, the non-polar organic solvent comprises an alkane solvent, an aromatic solvent, or a combination thereof. The alkane solvent can have 6 to 20 carbons. For example, an alkane can have 6 to 14 carbons, 6 to 9 carbons, and the like. Alternatively, alkanes can have 10-14 carbons. In certain embodiments, the alkane is a linear alkane. For example, the alkane solvent may include pentane, hexane, heptane, octane, decane, undecane, dodecane, or a combination thereof. In another embodiment, the alkane is halogenated. An exemplary branched alkane may contain a halogenated dimer of C11 or C12 alpha olefin.
In a further embodiment, the organic solvent may include a polar aprotic solvent. For example, polar aprotic solvents can include tetrahydrofuran, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide, N-methylpyrrolidone (NMP), or a combination thereof. In another example, the organic solvent may be free of ether solvents and may include, for example, dimethylformamide, acetonitrile, dimethyl sulfoxide, or a combination thereof.
As shown in 804, the wash solution and sensor device can be heated while the sensor pad is exposed to the wash solution. In one embodiment, the wash solution and sensor device can be heated at temperatures in the range of 35 ° C to 60 ° C. For example, the temperature can be in the range of 40 ° C to 50 ° C.
As shown in 806, the sensor array can be exposed to a wash solution and optionally heated for a period ranging from 30 seconds to 30 minutes. For example, the period can range from 30 seconds to 10 minutes, from 30 seconds to 5 minutes, or even from 1 minute to 2 minutes.
As shown in 808, the sensor array can be rinsed with organic solvent or water after exposure to the wash solution. The organic solvent can be a low boiling organic solvent. An exemplary low boiling organic solvent may have a standard boiling point, such as a standard boiling point in the range of 25 ° C to 100 ° C, 50 ° C to 100 ° C. In one example, the low boiling organic solvent comprises alcohol, ethanol, propanol and the like. Alternatively, the organic solvent can be N-methylpyrrolidone (NMP). Alternatively or additionally, the sensor array may be rinsed with water, deionized water, etc. In certain embodiments, the organic solvent is at least partially miscible with the organic solvent of the water and acid / solvent mixture.
As shown in 810, the sensor array can be dried. In one embodiment, the sensor array can be dried in an inert atmosphere, such as with dry nitrogen or helium. In another embodiment, the sensor can be heated in a dry atmosphere. In a further embodiment, the sensor array can be dried under vacuum.
In a further embodiment shown in FIG. 9, method 900 comprises exposing the sensor array to a wash solution containing an acid as described above, eg, a sulfonic acid as shown in 902. The sensor device can include a sensor array and optionally the well structure can define a well array operably corresponding to the sensor array. Wells in the well array can expose the sensor pads of the sensors in the sensor array. The washing solution may have the composition described above. Optionally, the sensor array and wash solution can be heated as shown in 904. For example, the sensor array and cleaning solution can be heated to the temperatures mentioned above.
In a further embodiment, the sensor array can be exposed to a wash solution and optionally heated for a period ranging from 30 seconds to 30 minutes, as shown in 906. Alternatively, the sensor array can be exposed to the wash solution and optionally heated over the period described above.
After exposure to the wash solution, the sensor array can be rinsed with a solvent, eg, a low boiling solvent, as shown in 908. An exemplary low boiling point solvent is described above. Alternatively, the solvent can be N-methylpyrrolidone (NMP). The sensor array may be rinsed once or more than once with a low boiling solvent.
After rinsing with a low boiling organic solvent, the sensor array can be rinsed with water, deionized water, etc., as shown in 910. After rinsing with water, the sensor array can be dried as shown in 912. For example, the sensor array may be exposed to dry nitrogen gas.
Once dry, the cap can be applied across the sensor array, as shown in 914. For example, the cap can be attached to the sensor array or intervening well structures using an adhesive. Specifically, the cap includes a fluid port that communicates with the space defined between the cap and the well structure across the sensor array. Optionally, the method can be applied to a wafer before separating the wafer into an array and capping the individual arrays.
In a further exemplary embodiment shown in FIG. 10, Method 1000 comprises exposing the sensor array to a wash solution containing an acid as described above, eg, a sulfonic acid, as shown in 1002. The cleaning solution can further contain an organic solvent. Specifically, the sensor array can include a well structure that defines an array of wells operably corresponding to the array of sensors. The sensor pad of the sensor is exposed by the wells of the well array. The cap can be attached across the sensor array or well array. The cap includes a fluid port that communicates with the space defined between the cap and the well array or sensor array. The wash solution can be applied into the space through the fluid port. The composition of the washing solution can be as described above.
Optionally, as shown in 1004, the wash solution and sensor array can be heated. For example, the wash solution and sensor array can be heated at the temperatures mentioned above.
The sensor array can be exposed to the wash solution and optionally heated over the first period, as shown in 1006. The first period may have a range of 30 seconds to 30 minutes, a range of the above-mentioned period, and the like.
In addition, the basic solution can be applied to the sensor array over a second period, as shown in 1008. For example, the basic solution can be applied through the fluid port of the cap into the space defined between the cap and the well structure or sensor array. The basic solution has a pH of more than 7, a pH of more than 8, and even a pH of more than 9. The basic solution contains a strong base such as sodium hydroxide or potassium hydroxide, or tetraalkylammonium hydroxide such as tetramethylammonium hydroxide or tetraethylammonium hydroxide. This base has a concentration of 0.05M to 1.5M. For example, this base may have a concentration of 0.05M to 1.0M, 0.05M to 0.5M, or the like. The sensor array may be exposed to the basic solution for a period of 20 seconds to 15 minutes, a period of 20 seconds to 5 minutes, a period of 30 seconds to 2 minutes, and the like. Alternatively, the weak acid solution can be replaced with a basic solution.
The sensor array may be exposed to the wash solution, followed by repeated exposures such as k times to the basic solution r or weak acid. For example, k can be 1, 2, or 3, but is generally less than or equal to 10.
After exposure to wash and basic solutions, the sensor array can be rinsed with organic solvent or water, as shown in 1010. Exemplary low boiling organic solvents include those mentioned above. Alternatively, the organic solvent can be N-methylpyrrolidone (NMP). The sensor array may be rinsed at least once with a low boiling organic solvent, followed by rinsing with water. Optionally, the sensor array can be dried, such as by exposure to dry nitrogen, as shown in 1012.
Alternatively, the method of FIG. 10 can be applied to wafers containing multiple arrays. After the wafer has dried, the wafer can be separated into individual arrays and capped on the individual arrays.
Such processing of the sensor array has been shown to improve the filling of polymer beads that incorporate the analyte and secure the analyte closer to the sensor pad of the sensor array. Specifically, such treatment improves the filling of polymer beads containing the amplified polynucleotide analyte on it. For example, such processing methods improve the performance of sequence sequencing devices. Such processing of the sensor array has been shown to improve the sensor signal and signal-to-noise ratio (SNR), especially the FET-based sensor array.
For example, FIG. 11 shows an exemplary sequencing system improved by the processing methods described above. In the particular embodiment shown in FIG. 11, the polymer particles can be used as a support for polynucleotides during sequencing techniques. For example, such hydrophilic particles can immobilize polynucleotides for sequencing using fluorescent sequencing techniques. In another example, hydrophilic particles can immobilize multiple replicating polynucleotides for sequencing using ion sensing techniques. Alternatively, the treatment described above can improve the polymer matrix binding to the surface of the sensor array. The polymer matrix can capture analytes such as polynucleotides for sequencing.
In general, polymer particles can be treated to include biomolecules containing nucleosides, nucleotides, nucleic acids (oligonucleotides and polynucleotides), polypeptides, saccharides, polypeptides, lipids, or derivatives or analogs thereof. .. For example, polymer particles can bind or adhere to biomolecules. The end or any inner part of the biomolecule can be attached or attached to the polymer particles. Polymer particles can be attached or attached to biomolecules using cross-linking chemicals. Cross-linking chemicals include covalent or non-covalent bonds including ionic, hydrogen, affinity, dipole-to-bipolar, van der Waals, and hydrophobic bonds. Cross-linking chemicals include, for example, the avidin moiety and the biotin moiety; the antigen epitope and its antibody or immunoreactive fragment; the antibody and the hapten; the digoxigenin moiety and the anti-digoxigenin moiety; the fluorescein moiety and the anti-fluorescein moiety; Factors; nucleases and nucleotides; lectins and polysaccharides; steroids and steroid-binding proteins; active compounds and active compound receptors; hormones and hormone receptors; enzymes and substrates; immunoglobulins and proteins A; or oligonucleotides or polynucleotides and their counterparts Includes affinity between binding partners such as fluorescein.
As shown in FIG. 11, the plurality of polymer particles 1104 can be placed in solution together with the plurality of polynucleotides 1102. Multiple particles 1104 can be activated or prepared to bind polynucleotide 1102. For example, particle 1104 can contain oligonucleotides that are complementary to some of the polynucleotides of multiple polynucleotides 1102. In another example, the polymer particles 1104 can be modified with the target polynucleotide 1104 using techniques such as biotin-streptavidin binding.
In certain embodiments, hydrophilic particles and polynucleotides are susceptible to polymerase chain reaction (PCR) amplification or recombinase polymerase amplification (RPA). For example, dispersed phase droplets 1106 or 1108 can be formed as part of an emulsion and contain hydrophilic particles or polynucleotides. In one example, polynucleotide 1102 and hydrophilic particles 1104 are low on each other so that a single polynucleotide 1102 is more likely to be present in similar dispersed phase droplets as a single hydrophilic particle 1104. Provided in concentrations and low proportions. Other droplets, such as droplet 1108, can contain a single hydrophilic particle and cannot contain a polynucleotide. Each droplet 1106 or 1108 can contain an enzyme, nucleotide, salt, or polynucleotide to facilitate replication of the polynucleotide.
In certain embodiments, an enzyme, such as a polymerase, is present that binds to or is in close proximity to hydrophilic or hydrogel particles of dispersed phase droplets. In one example, the polymerase is present in the dispersed phase droplets to facilitate the replication of the polynucleotide. Various nucleic acid polymerases can be used in the methods described herein. In one exemplary embodiment, the polymerase can include an enzyme, a fragment or subunit thereof, which can catalyze the replication of a polynucleotide. In another embodiment, the polymerase is a naturally occurring polymerase, recombinant polymerase, mutant polymerase, variant polymerase, fusion, or otherwise modified polymerase, chemically modified polymerase, synthetic molecule, or analog. , Derivatives, or fragments thereof.
Following PCR or RPA, particles, particles 1110, etc. are formed, which can contain multiple replicas of hydrophilic particles 1112 and polynucleotide 1114. Polynucleotides 1114 are shown to be present on the surface of particle 1110, but polynucleotides can extend within particle 1110. Hydrogels and hydrophilic particles have a low concentration of polymer with respect to water and can be contained within and throughout the polynucleotide segment, which can be present in pores and other openings. Specifically, particle 1110 can be diffused by a polymer matrix with enzymes, nucleotides, primers, and reaction products used to observe the reaction. The large number of polynucleotides per particle produces a better signal.
In embodiments, the polymer particles from the emulsion breaking procedure can be recovered and washed in preparation for sequencing. Recovery can be done by contacting the biotin moiety with the avidin moiety (eg, connecting it to an amplified polynucleotide template that adheres to the polymer particles) and separating the biotinylated template from the polymer particles lacking. Recovered polymer particles carrying double-stranded template polynucleotides can be modified to provide single-stranded template polynucleotides for sequencing. The denaturation step can include treatment with a base (eg, NaOH), formamide, or pyrrolidone.
In one exemplary embodiment, particle 1110 can be used in a sequencing device. For example, the sequencer 1116 can include an array of wells 1118. The sequencing device 1116 can be treated with the wash solution containing the above-mentioned sulfonic acid. Particle 1110 can be placed in well 1118.
In one example, the primer can be added to well 1118 and the particles 1110 can be pre-exposed to the primer prior to placement in well 1118. Specifically, particle 1110 can include binding primers. Primers and polynucleotides form a nucleic acid double strand containing a polynucleotide (eg, a template nucleic acid) hybridized with the primer. Nucleic acid duplexes are at least partially duplex polynucleotides. Enzymes and nucleotides may be provided in well 1118 to facilitate detection reactions, eg, nucleotide integration.
Sequencing can be done by detecting the addition of nucleotides. For example, the addition of nucleotides can be detected by using a method such as a fluorescence emission method or an ion detection method. For example, a set of fluorescently labeled nucleotides can be provided to system 1116 and transferred to well 1118. Excitation energy can also be provided to wells 1118. When a nucleotide is captured by a polymerase and the nucleotide is added to the end of an extending primer, the nucleotide label can fluoresce to indicate what type of nucleotide was added.
In an alternative embodiment, the solution comprises a single type of nucleotide and can be fed sequentially. The pH of well 1118 in the local environment can change in response to nucleotide additions. Such changes in pH can be detected by an ion-sensitive field effect transistor (ISFET). Therefore, changes in pH can be utilized to generate a signal indicating the order of nucleotides complementary to the polynucleotide of particle 1110.
Specifically, the sequencing system may include placing wells or wells on sensor pads such as ion sensors, such as field effect transistors (FETs). In an embodiment, the system is placed on one or more polymer particles filled in wells placed on the sensor pad of an ion sensor (eg, FET), or on the sensor pad of an ion sensor (eg, FET). Contains one or more polymer particles packed in multiple wells. In embodiments, the FET can be a chemFET or an ISFET. A "chemFET", or chemical field effect transistor, includes a type of field effect transistor that acts as a chemical sensor. A chemFET has a structural analog of a MOSFET transistor, in which a change in the gate electrode is applied by a chemical process. An "ISFET", i.e., an ion-sensitive electric field effect transistor, can be used to measure the ion concentration in solution, and when the ion concentration (eg, H +) changes, the current in the transistor changes accordingly.
In the embodiment, the FET may be a FET array. As used herein , an "array" is a planar array of elements, such as a sensor or well. The array may be one-dimensional or two-dimensional. A one-dimensional array may be an array having one stage (or column) of elements in the first dimension and multiple stages (or columns) of components in the second dimension. The number of columns (or columns) in the first and second dimensions may or may not be the same. FET or array is 10<sup>2</sup>、10<sup>3</sup>、10<sup>4</sup>、10<sup>5</sup>、10<sup>6</sup>、10<sup>7</sup>Or more FETs can be included.
In embodiments, one or more microfluidic structures can be assembled on FET sensor arrays to provide containment or confinement of biological or chemical reactions. For example, in one implementation, microfluidic structures (s) are placed on one or more sensors in an array so that one or more wells (or terms are used interchangeably herein). Can be configured as a microwell, or reaction chamber, or reaction well), whereby one or more sensors on which a given well is placed will have the presence of an analyte in the given well. Detect or measure level or concentration. In the embodiment, there may be a 1: 1 correspondence between the FET center and the reaction well.
Returning to FIG. 11, in another embodiment, wells 1118 in an array of wells can be operably connected to the measurement device. For example, in the fluorescence emission method, well 1118 can be operably linked to a photodetector. For ion detection, the lower surface of well 1118 can be placed on an ion sensor, such as a sensor pad such as a field effect transistor.
An exemplary system that includes sequencing by detection of nucleotide-incorporated ion by-products is the Ion Torrent PGM or Proton Sequencer (Life Technologies), which are ionic sequencing systems. Nucleic acid templates are sequenced by detecting hydrogen ions produced as by-products of nucleotide integration. Typically, hydrogen ions are released as a by-product of nucleotide integration that occurs during the synthesis of template-dependent nucleic acids by polymerases. The Ion Torrent PGM or Proton sequencer detects nucleotide integration by detecting hydrogen ion by-products of nucleotide integration. Ion Torrent A PGM or Proton sequencer can include multiple template polynucleotides, the template polynucleotides being sequenced, and each template placed in its own sequencing reaction well within the array. Will be done. Each well of the array can be connected to at least one ion sensor, which can detect the release of H + ions or changes in the pH of the solution produced as a by-product of nucleotide integration. .. The ion sensor includes a field effect transistor (FET), which is connected to an ion sensitive detection layer, which can detect the presence of H + ions or changes in the pH of the solution. The ion sensor can provide an output signal indicating nucleotide integration, which can be indicated as a change in voltage, the magnitude of which change correlates with the H + ion concentration in each well or reaction chamber. To do. Multiple types of nucleotides can be continuously introduced into the reaction chamber and incorporated into the extending primers (or polymerization sites) using a polymerase in the order determined by the sequence of the template. Each nucleotide integration can be achieved by releasing H + ions in the reaction well and at the same time varying the local pH. The FET of the sensor can record the emission of H + ions, which produces a signal indicating the occurrence of nucleotide integration. Nucleotides that are not incorporated into a particular nucleotide stream cannot generate a signal. The amplitude of the signal from the FET can also correlate with the number of nucleotides of a particular type, which are incorporated into the extending nucleic acid molecule, thereby dividing the homopolymer region. Thus, during sequencer operation, multiple nucleotides flow into the reaction chamber and at the same time are observed incorporated into various wells, or the reaction chamber allows the instrument to simultaneously split the sequences of multiple nucleic acid templates. It becomes possible to do.
In the first aspect, the sensor array comprises a plurality of sensors. One of the sensors has a sensor pad that is exposed to the surface of the sensor array. The method of processing the sensor array includes at least exposing the sensor pad to a cleaning solution containing an acid and an organic solvent and rinsing the cleaning solution from the sensor pad.
In the second aspect, the sensor array comprises a plurality of sensors. One of the sensors includes a sensor pad. The well structure defines a well array that corresponds to the sensor array. Wells in the well array expose the sensor pad. The cap is mounted over the sensor array and well structure and includes a fluid port. The space is defined between the cap and the well structure. Methods of processing the sensor array include applying it into space through a fluid port and waiting for a first period of 30 seconds to 30 minutes. The cleaning solution contains an acid and an organic solvent. The method further comprises applying the basic solution into the space through the fluid port and waiting for a second period of 20 seconds to 15 minutes, and applying the rinse solution through the fluid port. In one embodiment of the second aspect, the method further comprises providing a wash solution and repeating the application of a basic solution.
In a third aspect, the sensor array comprises a plurality of sensors. One of the sensors includes a sensor pad. The well structure defines a well array that corresponds to the sensor array. Wells in the well array expose the sensor pad. The method of processing the sensor array includes providing the cleaning solution to at least the sensor pad and waiting for a first period of 30 seconds to 30 minutes. The cleaning solution contains an acid and an organic solvent. The method further includes at least rinsing the sensor pad with a low boiling organic solvent and drying the sensor array. In the embodiment of the third aspect, the method further comprises rinsing with water after rinsing with a low boiling organic solvent and before drying. In another embodiment and the above embodiment of the third aspect, the method further comprises attaching the cap to the sensor array and the well array, the cap comprising a fluid port, and the space being the cap and the well array. It is defined between and communicates with the fluid port.
In one and above embodiments of the first, second, and third aspects, the acid comprises a sulfonic acid. The sulfonic acid can include alkyl sulfonic acid, alkylaryl sulfonic acid, or a combination thereof. In one example, the alkylaryl sulfonic acid comprises an alkyl group having 1 to 20 carbons. For example, an alkyl group has 9 to 18 carbons, 10 to 14 carbons, and the like. In another example, the alkyl group has 1 to 6 carbons.
In another embodiment and the above embodiment of the first, second, and third aspects, the sulfonic acid comprises dodecylbenzenesulfonic acid. In further and above embodiments of the first, second, and third embodiments, the sulfonic acid comprises methanesulfonic acid, ethanesulfonic acid, propanesulfonic acid, butanesulfonic acid, or a combination thereof. In further and above embodiments of the first, second, and third embodiments, the sulfonic acid comprises paratoluenesulfonic acid.
In further and above embodiments of the first, second, and third aspects, the wash solution comprises 10 mM to 500 mM acid. For example, the cleaning solution contains 50 mM to 250 mM acid.
In further and above embodiments of the first, second, and third aspects, the wash solution comprises 0.5% to 25% by weight of acid. For example, the cleaning solution contains 1% by weight to 10% by weight of acid, 2.5% by weight to 5% by weight of acid, and the like.
In another embodiment and the above embodiment of the first, second, and third aspects, the organic solvent is non-polar.
In further and above embodiments of the first, second, and third embodiments, the organic solvent has a standard boiling point in the range of 36 ° C to 345 ° C. For example, the standard boiling point is in the range of 65 ° C to 275 ° C. In one example, the standard boiling point is in the range of 65 ° C to 150 ° C. In alternative embodiments, the standard boiling point is in the range of 150 ° C to 220 ° C.
In further and above embodiments of the first, second, and third aspects, the organic solvent is an alkane having 6 to 24 carbons. For example, alkanes have 6-14 carbons, 6-9 carbons, or 10-14 carbons, etc.
In further and above embodiments of the first, second, and third aspects, the organic solvent is an aprotic polar solvent. For example, polar aprotic solvents include tetrahydrofuran, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide, N-methylpyrrolidone (NMP), or a combination thereof.
In another embodiment and the above embodiment of the first, second, and third aspects, the method further comprises heating the sensor pad and the cleaning solution while exposing the sensor pad to the cleaning solution. For example, heating involves heating at a temperature in the range of 35 ° C to 70 ° C. For example, the temperature is in the range of 40 ° C to 55 ° C.
In a further embodiment and above of the first, second, and third embodiments, exposure involves exposure for a period ranging from 30 seconds to 30 minutes. For example, the period may be in the range of 30 seconds to 10 minutes, in the range of 30 seconds to 5 minutes, or in the range of 1 minute to 3 minutes.
In further and above embodiments of the first, second, and third aspects, the method further comprises exposing at least the sensor pad to a basic solution. For example, the basic solution contains 0.005M to 1.5M sodium hydroxide, 0.01M to 1.0M sodium hydroxide, 0.01M to 0.5M sodium hydroxide, and the like. In a further embodiment, at least the exposure of the sensor pad to the basic solution occurs after the sensor pad is exposed to the wash solution. The method can further include at least exposing the sensor pad to a wash solution and at least repeatedly exposing the sensor pad to a basic solution.
In another embodiment and the above embodiment of the first, second, and third embodiments, rinsing involves rinsing with a low boiling organic solvent having a standard boiling point in the range of 25 ° C to 100 ° C. .. For example, low boiling organic solvents have a standard boiling point in the range of 50 ° C to 100 ° C. In certain embodiments, the low boiling organic solvent comprises alcohol. For example, alcohol includes ethanol or isopropanol.
In further and above embodiments of the first, second, and third aspects, rinsing involves rinsing with water.
In a fourth aspect, the system comprises a sensor device that is processed by any one of the above aspects and embodiments.
In a fifth aspect, the solution comprises 2.5% to 5% by weight of sulfonic acid and a non-polar linear alkane having 6 to 8 carbons.
In a sixth aspect, the kit comprises a nucleotide solution containing at least one nucleotide type, polymer particles, and a wash solution containing 0.5% to 20% by weight of sulfonic acid, and a standard in the range of 65 ° C to 275 ° C. Contains an organic solvent having a boiling point.
Not all of the above-mentioned activities described in the outline or examples are required, some of the specific activities may not be required, and one or more activities may be further performed in addition to the above-mentioned activities. Please note that there is. Still further, the order in which the activities are described is not necessarily the order in which the activities take place.
Specific salts of the acids or bases mentioned above can be useful in the methods described above. Salts include alkaline or alkaline earth metal salts, or tetraalkylammonium salts of acids or bases above.
The above specification describes the concept with reference to a particular embodiment. However, one of ordinary skill in the art will appreciate that various modifications and modifications can be made without departing from the scope of the invention, as described in the claims below. Therefore, the specification and drawings should be viewed in an exemplary sense rather than in a limiting sense, and all such modifications are intended to be included within the scope of the invention.
As used herein, the terms "comprises", "comprising", "includes", "including", "has", "has", "includes", "includes", "includes", "includes" "Having", or other variants thereof, is intended to cover non-exclusive inclusion. For example, a process, method, article, or device contains a list of features, but is not necessarily limited to those features and is not explicitly described, or is specific to such process, method, article, or device. Other features can be included. Furthermore, on the contrary, unless explicitly stated, "or" refers to an inclusive "or" rather than an exclusive "or". For example, condition A or B is satisfied by any one of the following: A is correct (or exists) and B is incorrect (or does not exist), A is incorrect (or does not exist) and B is correct (or exists), and both A and B are correct (or exist).
Also, adopting the use of "one (a)" or "one (an)" to describe the elements or components described herein. This is done solely for convenience and provides general implications for the scope of the invention. Unless otherwise clear, those statements should be construed to include one or at least one, and the singular form also includes plurals.
The benefits, other benefits, and solutions to the problems are described above with reference to embodiments. However, any benefit, benefit, solution to a problem, and any feature (s) may result in or manifest any benefit, benefit, or solution, but any or all claims. It is not to be interpreted as an important, essential, or essential feature of.
Those skilled in the art will appreciate that certain features may be described herein to be articulated in the context of individual embodiments and provided in combination within a single embodiment. Will be understood after interpreting. Conversely, various features are described for brevity in the context of a single embodiment, but may be provided individually or in any partial combination. Furthermore, references to the values listed in the range include each and all values within the range.
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Numbers
- Publication
- 6450366
- Publication, DOCDB
- 6450366
- Publication, EPODOC
- JP6450366B
- Application
- 2016505602
- Application, DOCDB
- 2016505602
- Application, EPODOC
- JP20160505602
Titles2
- Japanese
- 半導体装置を処理するための方法
- English
- Methods for processing semiconductor devices
Classification
- CPC, 10
- G01N27/4145
- B08B9/08
- C11D3/162
- C11D3/3418
- C11D3/43
- C11D7/5027
- B08B3/08
- C11D7/34
- C11D7/5013
- C11D7/5022
- IPC, 1
- G01N27 414
