Buffers and their applications

A technology of buffer and acetate buffer, which is applied in the field of joint connection buffer, buffer, and construction of nucleic acid library, which can solve the limitations of high-throughput workflow construction, limited height of the connection reaction plateau, and inability to advance the connection time, etc. problems, to achieve the effect of improving the efficiency of the ligation reaction, reducing the amount of enzyme used in the system, and having a small inhibitory effect
CN111100905BActive Publication Date: 2021-04-06ZHEJIANG ANNOROAD BIO TECH CO LTD +1
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Patent Information

Authority / Receiving Office
CN Β· China
Patent Type
Patents(China)
Current Assignee / Owner
ZHEJIANG ANNOROAD BIO TECH CO LTD
Publication Date
2021-04-06

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Abstract

The invention discloses a buffer and an application thereof, wherein the buffer comprises: 33-66mM Tris buffer, 1.6-5mM divalent cations, 25-75mM monovalent cations, 0.5-10mM dithiothreitol (DTT), 5‑15% PEG4000‑8000 and 0.5‑2mM ATP, pH 7‑9. The buffer can improve the efficiency of the ligation reaction, increase the conversion rate of the substrate, reduce the mismatch rate of the ligation reaction, and has good compatibility and little inhibitory effect on the ligase. At the same time, it can significantly reduce the amount of enzyme used in the system, thereby reducing the experimental the cost of.
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Description

technical field

[0001] The present invention relates to the field of biotechnology, in particular to a buffer and its application, more specifically to a buffer, a kit, the buffer used in the construction of a nucleic acid library as an adapter ligation buffer, and a Methods for constructing nucleic acid libraries. Background technique

[0002] The general process for constructing a next-generation sequencing library includes: (1) Fragmenting the target DNA; (2) Blunting the free DNA; (3) Protruding adenylation at the 3' end of the blunt DNA ; (2) Ligate the protruding adenylated DNA fragment with the protruding thymine-prominent double-stranded Y-shaped linker. The purpose of library construction is to add adapters to the two segments of double-stranded DNA fragments. The library conversion rate is the ratio of the measured yield to the theoretical maximum yield quality check, that is, how many initial samples are finally converted into two fragments with adapters. [00...

Claims

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