Construction method of single cell transcriptome sequencing library and application of construction method

Abstract

The invention discloses a construction method of a single cell transcriptome sequencing library and application of the construction method. The construction method includes the following steps of preparing a cDNA sample from a single cell and conducting fragmentized library construction on the cDNA sample to obtain the transcriptome sequencing library of the single cell, wherein fragmentation is conducted on the cDNA sample with a physical breaking method in the step of constructing the fragmentized library, and the step for purifying the fragmentized cDNA sample is not needed. According to the method, fragmentation is conducted on the sample of the single cell with the physical breaking method, and therefore fragments obtained through breaking are small and relatively uniform, and the peak width of the constructed single cell transcriptome sequencing library is concentrated; the purification step is not needed for the fragmentized sample, sample loss is reduced, construction of the single cell transcriptome sequencing library of the micro-sample can be achieved, stability of output of data volume can be improved, and therefore the reliability of large-scale and high-throughput sequencing of the single cell transcriptome sequencing library is improved.

Description

technical field [0001] The invention relates to the technical field of high-throughput sequencing, in particular to a method for constructing a single-cell transcriptome sequencing library and its application. Background technique [0002] With the development of modern biology, the study of cell populations can no longer meet the needs of scientific research. Single-cell research can not only solve the problem of cell heterogeneity that cannot be solved by tissue cells, but also provide a basis for analyzing the behavior, mechanism and The relationship with the organism provides a new research direction. At present, single-cell sequencing, which applies high-throughput sequencing to single-cell research, has become a new way of single-cell research, mainly including single-cell whole-genome sequencing and single-cell transcriptome sequencing. [0003] For single-cell transcriptome sequencing, the construction of transcriptome sequencing library is the main difficulty, beca...

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Problems solved by technology

Because if the data output is too much, it will waste costs; if the output is insufficient, additional testing will be required later, which will delay the project cycle

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