Buffer solution composition and application thereof

Abstract

The invention discloses a buffer solution composition and application thereof. The buffer solution composition comprises: a general buffer solution, wherein the general buffer solution comprises 5-25 mMol of a general Tris buffer solution, 5-20 mMol of general divalent cations, 75-150 mMol of general monovalent cations, 0.05-2% by mass of a surfactant and 0.5-2 mMol of dithiothreitol (DTT), and the pH value of the general buffer solution is 8-10; and a connection buffer solution, which comprises 33 to 66 mM of a connection Tris buffer solution, 0.5 to 5 mM of connection divalent cations, 0.5 to 10 mM of dithiothreitol (DTT), 5 to 15% of PEG-4000 to PEG-8000 and 0.5 to 2 mM of ATP and has a pH value of 7 to 9. Therefore, the buffer solution composition can be used for library construction of multiple samples, and is short in reaction time, high in conversion rate and low in mismatch rate.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular, to a buffer composition and its application, more specifically, to a buffer composition, a kit, and a method for constructing a nucleic acid library. Background technique [0002] Next-generation sequencing of genomic DNA and corresponding bioinformatics analysis have been widely used in the fields of medical health and scientific and technological services. [0003] The general process for the construction of a next-generation sequencing library is as follows: fragment the target DNA; blunt the fragmented DNA; protrude adenylation at the 3' end of the blunt DNA; The DNA fragments are ligated with double-stranded Y adapters that highlight thymine. The existing buffer system has poor sample compatibility and is also difficult to be compatible with different library construction steps. Frequent buffer replacement is required, which prolongs the time for library construction, and...

AI Technical Summary

Problems solved by technology

The conversion rate is largely affected by the ligation efficiency. The ligation reaction efficiency is related to many factors such as the enzyme reaction efficiency, the openness of the DNA chain end, the collision efficiency of the donor and the acceptor, and the linker in the ligation reaction system will also The effective number of adapters generated in the self-ligating depletion reaction is an efficiency bottleneck in the library construction process

At present, the common rapid connection buffer in the market has a limited connection reaction plateau height, and there are situations such as increasing the amount of enzyme input to overcome the mismatch of the buffer system, which increases the cost of practical application; in addition, the connection time under the existing connection buffer Unable to push forward the enzyme reaction balance; and a large number of mismatches will be generated during the premixing process, which reduces the library yield and limits the construction of high-throughput workflows

[0004] Therefore, the general buffer and connection buffer in the existing library construction need to be improved

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