Cot DNA suitable for bisulfite sequencing and preparation method and application thereof

Abstract

The invention discloses Cot DNA suitable for bisulfite sequencing as well as a preparation method and application thereof. The method comprises the following steps of obtaining fragmented genome DNA, obtaining a transformed genome DNA fragment, obtaining a connection product, and obtaining Cot DNA, wherein the connection product is obtained by connecting the transformed genome DNA fragment with a joint and then amplifying, and the Cot DNA is obtained by performing denaturation, renaturation and purification treatment on the connection product. The Cot DNA provided by the invention is competitively hybridized with the repetitive sequence in the target library, so that the influence of the repetitive sequence on the captured nucleic acid is reduced, and the hybridization efficiency and target rate of the capture probe are improved.

Description

technical field [0001] The invention relates to the technical field of bisulfite sequencing library, in particular to CotDNA suitable for bisulfite sequencing library, its preparation method and application. Background technique [0002] In molecular biology, Cot is a unit in DNA renaturation kinetics, which represents the renaturation time (t, sec) of DNA single strand at a unit concentration (Co, mol / L), and the Cot value essentially represents The product of the initial single-strand concentration (Co) in the hybridization solution and the reaction time (t). Under the same conditions, DNA molecules with repetitive sequences have small Cot values, while DNA molecules with non-repeated sequences have larger Cot values. Cot DNA is usually used as a DNA competitive blocker to block repetitive sequences that may cause non-specific hybridization in genomic hybridization. For example, when probes containing repetitive sequences are used for experiments such as Northern hybridi...

AI Technical Summary

Problems solved by technology

However, in the DNA library established after the sample is processed, a large number of bases have changed compared with the original sequence, and the sequence structure of the repetitive sequence (especially in the animal genome, such as the human genome, there are a large number of repetitive sequences) has also changed. Can adversely affect subsequent targeted capture

Although repetitive sequences exist widely, their biological significance has not yet been clarified, and studies on methylation modification in epigenetics mainly focus on the promoter region and the first exon region, so a method that can detect repetitive sequences is needed. Blocking substances to reduce their impact on methylation detection results

[0005] At present, there is no Cot DNA suitable for bisulfite sequencing. The existing Cot DNA used to block these repetitive sequences in the sequence cannot adapt to the bisulfite sequencing library with base changes, and cannot achieve a good effect of blocking repetitive sequences.

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