A method for constructing a plasmid containing tandem repeat sequences

CN119776394APending Publication Date: 2025-04-08SHENZHEN RHEGEN BIOTECHNOLOGY CO LTD +1
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Patent Information

Application Number
CN202311301708.6
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2023-10-07
Publication Date
2025-04-08

AI Technical Summary

Technical Problem

In the prior art, when constructing plasmids containing long fragments tandem repeats, the problem of base increases or deletion is prone to occur, resulting in complex, time-consuming and low success rate.

Method used

By providing the first and second nucleic acid molecules, each with tandem repeats and annealing under gradient temperature conditions, the complementary sequences are bound to form a double-stranded nucleic acid fragment with tandem repeats, which is then attached to the carrier backbone with a sticky end.

Benefits of technology

The efficiency of correct pairing between long fragment tandem repeat primers is significantly improved, and the success rate of connecting double-stranded nucleic acid fragments containing tandem repeat sequences to the vector backbone is improved, more positive clones are obtained, and construction time is reduced.

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Abstract

The present invention provides a method for constructing a plasmid containing a tandem repeat sequence, comprising: providing a first upstream primer and a first downstream primer; the first upstream primer and the first downstream primer respectively have a tandem repeat sequence, and the tandem repeat sequence of the first upstream primer is complementary to at least part of the tandem repeat sequence in the tandem repeat sequence of the first downstream primer; annealing the first upstream primer and the first downstream primer under a gradient temperature condition to obtain a first double-stranded nucleic acid fragment having a tandem repeat sequence, wherein the tandem repeat sequence complementary to each other in the double strand is the first double-stranded tandem repeat sequence; connecting the first double-stranded nucleic acid fragment to a first vector backbone with a sticky end to construct a plasmid containing the first double-stranded tandem repeat sequence. The present invention can significantly improve the success efficiency of tandem repeat sequence connection, obtain more positive clones, ensure the success rate of the experiment, and reduce the construction time.
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Citation Information

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