Library preparation method, kit and sequencing method

A library and sequencing technology, applied in the biological field, can solve problems that need to be improved

Pending Publication Date: 2021-09-03
GENEMIND BIOSCIENCES CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Therefore, the existing library construction method and sequencing method ad

Method used

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  • Library preparation method, kit and sequencing method
  • Library preparation method, kit and sequencing method
  • Library preparation method, kit and sequencing method

Examples

Experimental program
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Effect test

Embodiment

[0076] The total RNA of empty-loaded E. coli DH5a was extracted as the initial nucleic acid, and the initial amount of sample nucleic acid was used in three gradients: 10pg, 50pg and 100pg, and the library construction effect of 6 kinds of magnetic beads was compared. At the same time, two sets of comparisons were set up for each magnetic bead In the experiment, three gradients of 10pg, 50pg, and 100pg were set for the initial amount of experimental samples in each group: the library construction method using magnetic beads enrichment and the library construction method not suitable for magnetic bead enrichment, and compared the effect of two different methods of library construction .

[0077] The same magnetic beads were used in the enrichment purification step and the magnetic bead purification step in the same library construction process, and the 6 kinds of magnetic beads used were: Group A Yeasen cfDNA magnetic beads (CatNO: 12599ES03, Yeasen), Group B Yeasen clean magnet...

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Abstract

The invention provides a library preparation method, a kit and a sequencing method. The library preparation method comprises the following steps: obtaining a target nucleic acid molecule, namely performing reverse transcription on RNA (Ribonucleic Acid) from a nucleic acid sample to be detected to obtain a reverse transcription product which comprises RNA: DNA (Deoxyribose Nucleic Acid) hybrid double strands and RNA: DNA hybrid double strands enriched in the reverse transcription product; providing a transposal complex, wherein the transposal complex comprises a linker and a transposase, the linker is a double-stranded nucleic acid molecule with a known sequence; contacting the target nucleic acid molecule and the transposition complex under conditions suitable for a transposition reaction to obtain a transposition product, wherein the transposition product comprises a double-stranded nucleic acid molecule with a joint at the tail end and a gap; providing a first primer configured to hybridize with at least a portion of the linker; and hybridizing the transposon with the first primer and placing under conditions suitable for strand displacement reaction to obtain the library. The method is particularly suitable for rapid construction of RNA libraries with low initial quantities.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular, the invention relates to a library preparation method, a library preparation kit and a sequencing method. Background technique [0002] The quality of sequencing data is related to the sequencing method and sequencing library. At present, many studies are devoted to the construction optimization of sequencing libraries and the optimization of sequencing methods. [0003] In the process of library preparation for trace amounts of nucleic acid (such as less than 1ng), the nucleic acid quality, preparation method, and reaction conditions used in the preparation process include inappropriate or abnormal solution systems, such as too low nucleic acid concentration, and the size of the screened nucleic acid fragment is different. Suitable, multiple components in the reaction system may interact and interfere with each other under specific conditions, which may lead to the preparation of low...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806C12N15/10C40B50/06C12Q1/6874G16B30/10
CPCG16B20/30G16B20/50G16B30/00G06N7/01
Inventor 林群婷甘广丽樊济才金欢张萌李改玲张娟刘丽春冯燕黄梦娴林玉琪
Owner GENEMIND BIOSCIENCES CO LTD
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