US 12,312,637 B2
Method for performing PCR reaction using comprehensive PCR reaction system
Qiaoling Han, Zhejiang (CN); Qiang Xu, Zhejiang (CN); and Xiangmin Cui, Zhejiang (CN)
Assigned to SHENYI BIOTECH (HANGZHOU) CO., LTD., Zhejiang (CN)
Filed by SHENYI BIOTECH (HANGZHOU) CO., LTD., Zhejiang (CN)
Filed on Dec. 14, 2021, as Appl. No. 17/551,153.
Application 17/551,153 is a continuation of application No. PCT/CN2020/097142, filed on Jun. 19, 2020.
Claims priority of application No. 201910585250.9 (CN), filed on Jul. 1, 2019; application No. 201910591817.3 (CN), filed on Jul. 1, 2019; application No. 201910591847.4 (CN), filed on Jul. 1, 2019; application No. 201921011114.0 (CN), filed on Jul. 1, 2019; application No. 201910684842.6 (CN), filed on Jul. 26, 2019; application No. 201910953740.X (CN), filed on Oct. 9, 2019; application No. 201910953826.2 (CN), filed on Oct. 9, 2019; application No. 201910954105.3 (CN), filed on Oct. 9, 2019; application No. 201910954116.1 (CN), filed on Oct. 9, 2019; and application No. 201921682904.1 (CN), filed on Oct. 9, 2019.
Prior Publication US 2022/0106626 A1, Apr. 7, 2022
Int. Cl. C12Q 1/686 (2018.01); B01L 3/00 (2006.01); B01L 7/00 (2006.01)
CPC C12Q 1/686 (2013.01) [B01L 3/502715 (2013.01); B01L 3/502738 (2013.01); B01L 7/52 (2013.01); B01L 2200/0689 (2013.01); B01L 2200/16 (2013.01); B01L 2300/044 (2013.01); B01L 2300/0877 (2013.01); B01L 2400/0478 (2013.01); B01L 2400/0622 (2013.01); C12Q 2527/125 (2013.01); C12Q 2565/629 (2013.01)] 20 Claims
OG exemplary drawing
 
1. A method for performing a PCR reaction using a PCR reaction system, the PCR reaction system comprising:
a sample containing unit having a pyrolysis freeze-dried powder and a sample provided therein, the sample containing unit comprising a first liquid outlet/inlet;
a diluent containing unit having a diluent provided therein, the diluent containing unit comprising a diluent outlet;
a PCR reaction unit having a freeze-dried powder of a reverse transcriptase and PCR raw materials provided, the PCR reaction unit comprising a PCR reaction solution outlet and a pyrolyzed sample mixture inlet; and
a piston unit comprising an injection chamber and a piston, the injection chamber comprising a second liquid outlet/inlet,
wherein the second liquid outlet/inlet is connected to the first liquid outlet/inlet through a first pipeline, the second liquid outlet/inlet is connected to the diluent outlet through a second pipeline, the second liquid outlet/inlet is connected to the pyrolyzed sample mixture inlet through a third pipeline, and the PCR reaction solution outlet is connected to the diluent outlet through a fourth pipeline,
the method comprising:
subjecting the piston to a first movement processing, to allow a part of the diluent to enter the injection chamber, wherein the diluent is provided in the diluent containing unit;
subjecting the piston to a second movement processing, to subject the part of the diluent entering the injection chamber, the pyrolysis freeze-dried powder, and the sample to a first mixing processing, wherein the pyrolysis freeze-dried powder and the sample are provided in the sample containing unit, and the first mixing processing is performed in the sample containing unit;
subjecting a product of the first mixing processing to a pyrolysis processing, wherein the pyrolysis processing is performed in the sample containing unit;
subjecting the piston to a third movement processing, to allow a product of the pyrolysis processing to enter the injection chamber;
subjecting the piston to a fourth movement processing, to subject the product of the pyrolysis processing entering the injection chamber and the remaining part of the diluent to a second mixing processing, wherein the second mixing processing is performed in the diluent containing unit;
subjecting the piston to a fifth movement processing, to allow a product of the second mixing processing to enter the injection chamber;
subjecting the piston to a sixth movement processing, to subject the product of the second mixing processing entering the injection chamber and the freeze-dried powder of the reverse transcriptase and the PCR raw materials to a third mixing processing, wherein the freeze-dried powder of the reverse transcriptase and the PCR raw materials is provided in the PCR reaction unit, and the third mixing processing is performed in the PCR reaction unit; and
subjecting a product of the third mixing processing to a PCR temperature cycle amplification processing, wherein the PCR temperature cycle amplification processing is performed in the PCR reaction unit.