Hubei University

Jiakai Cui

Lixin Ma

Fei Wang

Wanping Chen

Longyu Wang

Rui Han

Xiaochen Xie

Nianzu He

Miaomiao Chen

PFU DNA POLYMERASE MUTANTS WITH REVERSE TRANSCRIPTASE ACTIVITY AND THEIR APPLICATIONS

An efficient synthesis and assembly method for the large fragment DNA based on the programmable nuclease Argonaute, specifically includes: constructing and treating antibiotic resistance gene reconstructed vectors with linearization, dividing a target DNA into multiple small DNA fragments and then synthesizing the small DNA fragments, followed by loading the synthesized small DNA fragments to the antibiotic resistance gene reconstructed vectors; and the SLIC and resistance gene reconstruction are used to achieve assembly of the target DNA. The method combines the SLIC with a resistance gene reconstruction strategy, allowing for the assembly of 5-6 small fragments in a single resistance gene reconstruction, which is more efficient and time-saving. Moreover, the number of the resistance gene reconstructions can be flexibly chosen according to the length of the DNA fragments. Mutations are not introduced caused by PCR, and the reconstructed large fragment do not need a second sequencing, saving time and costs.

Efficient synthesis and assembly method for large fragment DNA based on programmable nuclease Argonaute

Growing community of inventors

Wuhan, China

Jiakai Cui