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Title:
MICROWELL ARRAY CHIP-BASED SINGLE MOLECULE LIBRARY PCR AMPLIFICATION GENE SEQUENCING METHOD
Document Type and Number:
WIPO Patent Application WO/2019/161810
Kind Code:
A2
Abstract:
Provided is a microwell array chip-based single molecule library PCR amplification gene sequencing method, comprising: step one, a PCR amplification solution is added to a chip having a microwell array in a dropwise manner, so that microwells on the chip are all independent reaction spaces, wherein only one DNA sample fragment is fixed in a microwell; step two, PCR amplification is performed on the sealed microwell chip in a PCR apparatus; step three, after the amplification has finished, the double-stranded DNA within the microwells is denatured into single-stranded DNA; step four, by means of an annealing method, a sequencing primer S2 is combined with the DNA samples for sequencing in the microwells; step five, during sequencing, a deoxynucleoside triphosphate (dNTP) is added to the microwells according to sequences of G, C, A and T, and after successful pairing, an increase in the charge of a DNA sample or a release of hydrogen ions or pyrophosphate ions (PPI) causes a signal response of a sensor at the bottom of a microwell, and the response signal of the sensor is recorded and processed to obtain gene sequencing information of the sequenced DNA; step six, step five is repeated.

Inventors:
LIU, Yabao (7 Fenghuang Avenue, Building E 1F Zhangjiagan, Suzhou Jiangsu 4, 215614, CN)
YUE, Yaofei (7 Fenghuang Avenue, Building E 1F Zhangjiagan, Suzhou Jiangsu 4, 215614, CN)
ZHANG, Zhifeng (7 Fenghuang Avenue, Building E 1F Zhangjiagan, Suzhou Jiangsu 4, 215614, CN)
Application Number:
CN2019/082561
Publication Date:
August 29, 2019
Filing Date:
April 12, 2019
Export Citation:
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Assignee:
ZHANGJIAGANG ONE-CHIP BIOTECHNOLOGY CO., LTD. (7 Fenghuang Avenue, Building E 1F Zhangjiagan, Suzhou Jiangsu 4, 215614, CN)
Attorney, Agent or Firm:
TSINGYIHUA INTELLECTUAL PROPERTY LLC (Room 301 Trade Building, Zhaolanyuan Tsinghua University, Qinghuayuan, Haidian District, Beijing 4, 100084, CN)
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