US 11,739,319 B2
PCR primer pair and application thereof
Lin Yang, Shenzhen (CN); Guodong Huang, Shenzhen (CN); Ya Gao, Shenzhen (CN); Yanyan Zhang, Shenzhen (CN); Haiping Zhang, Shenzhen (CN); Fang Chen, Shenzhen (CN); Haojun Jiang, Shenzhen (CN); Shaohong Zhang, Shenzhen (CN); Yuqian Wang, Shenzhen (CN); Yifan Xie, Shenzhen (CN); and Hui Jiang, Shenzhen (CN)
Assigned to MGI TECH CO., LTD., Shenzhen (CN)
Appl. No. 16/624,779
Filed by MGI TECH CO., LTD., Shenzhen (CN)
PCT Filed Jun. 20, 2017, PCT No. PCT/CN2017/089196
§ 371(c)(1), (2) Date Dec. 19, 2019,
PCT Pub. No. WO2018/232595, PCT Pub. Date Dec. 27, 2018.
Prior Publication US 2020/0216838 A1, Jul. 9, 2020
Int. Cl. C12Q 1/68 (2018.01); C12P 19/34 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01)
CPC C12N 15/1093 (2013.01) [C12N 15/11 (2013.01)] 6 Claims
 
1. A PCR primer pair, comprising a first primer and a second primer, wherein
the first primer comprises a first specific sequence and a first random sequence, and
the second primer comprises a second specific sequence and a second random sequence, wherein
the first specific sequence is located at the 3′ end of the first primer and the first random sequence is located at the 5′ end of the first primer,
the second specific sequence is located at the 3′ end of the second primer and the second random sequence is located at the 5′ end of the second primer,
the first specific sequence and the second specific sequence are respectively an upstream primer and a downstream primer for a target sequence,
the first random sequence and the second random sequence are reversely complementary,
the first specific sequence is connected to the first random sequence via a first predetermined restriction enzyme cutting site,
the second specific sequence is connected to the second random sequence via a second predetermined restriction enzyme cutting site, and
the target sequence of the PCR primer pair does not comprise any predetermined restriction enzyme cutting site,
wherein the first random sequence and the second random sequence each have a length of 15-45 bp, and
the first specific sequence and the second specific sequence each have a length of 15-30 bp,
wherein the first predetermined restriction enzyme cutting site and the second predetermined restriction enzyme cutting site each are a uracil (U) base site.