Library building method and application

Abstract

The invention provides a library building method which comprises the following steps: (1) carrying out splitting decomposition treatment on a paraffin-embedded biological sample, and collecting a nucleic acid sample from a splitting decomposition solution obtained by the splitting decomposition treatment by utilizing a precipitation method, the nucleic acid sample containing a nucleic acid fragment with the length of less than 200bp; (2) carrying out terminal repair on the concentrated nucleic acid and adding a joint so as to obtain a nucleic acid sample connected with the joint; and (3) carrying out amplification treatment on the nucleic acid sample connected with the joint so as to obtain amplification products, and forming the sequencing library by the amplification products. The method is suitable for library building of trace and fragmented nucleic acid samples, the library building success rate can be effectively increased, the read repetition rate of subsequent informatics analysis is effectively reduced, and the genome comparison rate is increased.

Description

technical field [0001] The invention relates to the field of bioinformatics, in particular, to a library building method, a sequencing library, a sequencing method, and a sequencing result analysis method. Background technique [0002] At present, the library construction method based on high-throughput sequencing is as follows: first, the complete long-chain DNA is randomly broken into short fragments with a length of about 350 bp, and then adapters are connected to both ends of each short fragment, and then PCR amplification is performed. Increase the amplification of the library. After obtaining DNA samples, adding adapters and amplifying the samples, the samples need to be purified. The current purification method usually uses magnetic bead purification. The principle of magnetic bead purification is mainly to use reagents to dehydrate and precipitate DNA, which is then connected by hydroxyl groups on the surface of magnetic beads. Na + Adsorbed with DNA, and then thro...

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Problems solved by technology

[0003] However, the commonly used high-throughput sequencing library construction methods at this stage cannot be successfully used for library construction of long-term and highly degraded formalin-fixed and paraffin-embedded (FFPE) samples, because the samples Rare, trace, and severely fragmented, the current library construction methods cannot target samples with high nucleic acid fragments below 200bp. Even if the library is successfully constructed, the repetition rate of nucleic acid reads is too high and the gene The comparison rate is too low, which also makes the analysis results unreliable

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