US 11,718,872 B2
Method for obtaining single-cell mRNA sequence
Fei Fan, Shenzhen (CN); Xiaofang Cheng, Shenzhen (CN); Wenwei Zhang, Shenzhen (CN); Weimao Wang, Shenzhen (CN); Luman Cui, Shenzhen (CN); and Ou Wang, Shenzhen (CN)
Assigned to MGI TECH CO., LTD., Shenzhen (CN)
Filed by MGI TECH CO., LTD., Shenzhen (CN)
Filed on Jun. 23, 2020, as Appl. No. 16/909,366.
Application 16/909,366 is a continuation of application No. PCT/CN2018/124293, filed on Dec. 27, 2018.
Claims priority of application No. PCT/CN2017/119342 (WO), filed on Dec. 28, 2017.
Prior Publication US 2020/0318181 A1, Oct. 8, 2020
Int. Cl. C12Q 1/6869 (2018.01); C12Q 1/6818 (2018.01)
CPC C12Q 1/6869 (2013.01) [C12Q 1/6818 (2013.01); C12Q 2565/514 (2013.01)] 19 Claims
OG exemplary drawing
 
1. A method for obtaining full-length sequences of mRNA molecules of a single cell, comprising the steps of:
(a) utilizing a carrier with a cell tag to capture mRNA molecules of the single cell, followed by performing a reverse transcription reaction to obtain cDNA molecules with cell tags,
(b) utilizing a transposase complex and a carrier with a molecule tag to obtain cDNA fragments with molecule tags based on a virtual compartment, wherein the transposase complex inserts to the cDNA randomly, and then the cDNA inserted with the transposase complex is captured by the carrier with molecule tags forming one virtual compartment, to add the molecule tags to the cDNA;
(c) high-throughput sequencing the cDNA fragments with the molecule tags obtained in step (b); and
(d) assembling sequencing results obtained in step (c) to obtain full-length sequence of each mRNA molecule according to the molecule tag and obtaining full-length sequences of all the mRNA molecules in each single cell according to the cell tag,
wherein the carrier with a cell tag is a solid carrier carrying the cell tag,
the carrier with a molecule tag is a solid carrier carrying the molecule tag,
each cDNA molecule derived from one single cell has a same cell tag and the cDNA molecules derived from different single cells have different cell tags, and
all the cDNA fragments derived from one same cDNA molecule have a same molecule tag and the cDNA fragments derived from different cDNA molecules have different molecule tags.