Method for preparing new-generation sequencing connector with double chain molecular label

Abstract

The invention provides a method for preparing a new-generation sequencing connector with a double chain molecular label. The method comprises the following steps of 1, synthetizing the sequence of a connector, wherein the sequence of the connector contains a first sequence and a second sequence; the second sequence includes protection basic groups of restrictive endonuclease, double chain molecular labels of seven random basic groups, and enzyme cutting sites of restrictive endonuclease HphI; the first sequence and the second sequence have complementary sequence segments; 2, performing annealing on the sequence of the connector; uniformly mixing the first sequence and the second sequence; performing annealing in an annealing system according to an annealing procedure to form double chains;3, supplementing the double chain connector formed in the second step; 4, performing enzyme cutting and recovering the connector. The method for preparing the new-generation sequencing connector withthe double chain molecular label has the advantages that the connector contains the longer double chain molecular label; a better information base is provided for information analysis; the method isapplicable to various kinds of sequencing platforms.

Description

technical field [0001] The invention relates to a method for preparing a next-generation sequencing joint with a double-strand molecular label, belonging to the field of molecular biology. Background technique [0002] In the Illumina sequencing platform, adapters are a necessary element in next-generation sequencing library construction. Currently, official adapters include various types, and the sequences include p5 and p7 amplification primer binding sequences, read1 and read2 sequencing primer binding sequences, and sample labels. sequence, molecular tag sequence, etc., and has a 3'-dT tail for A / T ligation. [0003] At present, single-strand molecular tags are usually designed at the p7 end to add tags to target molecules in the library, so that the bioinformatics analysis process can capture more refined molecular information and reduce the background noise of variation analysis. [0004] In addition, double-strand molecular tags can also be combined with single-stran...

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Problems solved by technology

[0005] However, there is currently no better method for adding double-stranded tags to facilitate library construction

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