Buffer solution and application thereof

A buffer and acetate buffer technology, which is used in the field of linker ligation buffer, buffer, and nucleic acid library construction, can solve the problem of the limited height of the ligation reaction platform, the limitation of high-throughput workflow construction, and the inability to advance the connection time. To improve the efficiency of the ligation reaction, reduce the amount of enzyme used in the system, and achieve a small inhibitory effect
CN111100905AActive Publication Date: 2020-05-05ZHEJIANG ANNOROAD BIO TECH CO LTD +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
ZHEJIANG ANNOROAD BIO TECH CO LTD
Publication Date
2020-05-05

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Abstract

The invention discloses a buffer solution and application thereof. The buffer solution is prepared from a Tris buffer solution with a concentration of 33 to 66 mM, divalent cations with a concentration of 1.6 to 5 mM, monovalent cations with a concentration of 25 to 75 mM, dithiothreitol (DTT) with a concentration of 0.5 to 10 mM, PEG 4000 to 8000 with a concentration of 5 to 15% and ATP with a concentration of 0.5 to 2 mM. The pH value of the Tris buffer solution is 7 to 9. The buffer solution can improve the efficiency of a ligation reaction, increase the conversion rate of a substrate and reduce the mismatch rate of the ligation reaction, is good in compatibility and small in inhibition effect on ligase, and remarkably reduces the enzyme consumption of a system, so experiment cost is reduced.
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Description

Technical field

[0001] The present invention relates to the field of biotechnology, in particular, to buffers and their applications, and more specifically to a buffer, a kit, the buffer used in the construction of a nucleic acid library as a linker ligation buffer, and a Methods of constructing nucleic acid libraries. Background technique

[0002] The general process of constructing a second-generation sequencing library includes: (1) Fragmentation of the target DNA; (2) End-blending treatment of free DNA; (3) Protruding adenylation at the 3'end of the blunted DNA (2) Connect the overhanging adenylated DNA fragment with the overhanging thymidine double-stranded Y-linker. The purpose of library construction is to add linkers to the two double-stranded DNA fragments. The library conversion rate is the ratio of the measured yield to the theoretical maximum yield quality inspection, that is, how many starting samples are finally converted into two fragments with adapters.

[0003] ...

Claims

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