US 12,077,814 B2
Method for low frequency somatic cell mutation identification and quantification
Liping Wei, Beijing (CN); Boxun Zhao, Beijing (CN); Yue Huang, Beijing (CN); Qixi Wu, Beijing (CN); Yongxin Ye, Beijing (CN); and Xianing Zheng, Beijing (CN)
Assigned to PEKING UNIVERSITY, Beijing (CN)
Appl. No. 16/616,865
Filed by PEKING UNIVERSITY, Beijing (CN)
PCT Filed Jul. 23, 2018, PCT No. PCT/CN2018/096620
§ 371(c)(1), (2) Date Nov. 25, 2019,
PCT Pub. No. WO2018/214989, PCT Pub. Date Nov. 29, 2018.
Claims priority of application No. 201710381726.8 (CN), filed on May 25, 2017.
Prior Publication US 2023/0265496 A1, Aug. 24, 2023
Int. Cl. C12Q 1/6869 (2018.01); C12N 15/10 (2006.01); C12Q 1/6883 (2018.01); C12Q 1/6886 (2018.01)
CPC C12Q 1/6869 (2013.01) [C12N 15/1093 (2013.01); C12Q 1/6883 (2013.01); C12Q 1/6886 (2013.01); C12Q 2600/118 (2013.01); C12Q 2600/156 (2013.01)] 19 Claims
OG exemplary drawing
 
1. A method of detecting genomic location of a transposon in a subject, comprising
a) providing isolated DNA sample from a test sample of the subject;
b) fragmenting the DNA sample to obtain DNA fragments;
c) ligating adaptors to both ends of the DNA fragments to obtain a transposon-adaptor library;
d) amplifying the transposon-adaptor library with primers comprising a first primer and a second primer, wherein the first primer comprises a transposon specific sequence capable of hybridizing to an internal sequence of the transposon, a second primer comprises an adapting sequence capable of hybridizing to the adaptor, thereby amplifying the transposon and flanking sequences thereof;
e) sequencing the obtained product from amplification to obtain the flanking sequences; and
f) aligning the obtained flanking sequences with a reference genomic sequence to obtain genomic location and orientation of the transposon; and
wherein the first primer is a primer set comprising 3 or more different first primers, wherein each of the first primer further comprises a library adapting sequence at its 5′ end and a spacer region is provided between the library adapting sequence and the transposon sequence, and wherein the length of the spacer region in each first primer is different.