Method for preparing next-generation sequencing linker with molecular tags

Abstract

The invention discloses a method for preparing a next-generation sequencing linker with molecular tags. The method is characterized by comprising the steps that 1, linker sequences are synthesized, wherein the linker sequences include a first sequence and a second sequence, the second sequence comprises protection bases and a double-chain molecular tag of six random bases of a restriction enzyme,and restriction enzyme cutting sites of the restriction enzyme MnlI, and the first sequence and the second sequence have complementary sequence segments; 2, the linker sequences are annealed, specifically, the first sequence and the second sequence are mixed evenly, and annealing is carried out in an annealing system according to an annealing process to form double chains; 3, an linker of the double chains formed in the step 2 is completed; and 4, restriction enzyme digestion is carried out, and the linker is recycled. The method for preparing the next-generation sequencing linker with the molecular tags, which is provided by the invention, has the following advantages that the linker has the longer double-chain molecular tag, so that a better information base is provided for bioinformation analysis; and the method adapts to various sequencing platforms.

Description

technical field [0001] The invention relates to a method for preparing a next-generation sequencing joint with molecular tags, which belongs 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-strand molecular t...

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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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