Transcriptome sequencing method

Abstract

The present invention provides a transcriptome sequencing method, wherein primers used during a reverse transcription process from RNA to cDNA comprise a random sequence and a specific sequence, the random sequence is positioned at the 3' terminal of the specific sequence, bases from 0 to 5 exist between the random sequence and the specific sequence, the random sequence comprises 4-12 random bases, the specific sequence is a homopolymer of 5-30 bases, and the base of the specific sequence is any one selected from A, G, C and T. According to the present invention, with the design on the reverse transcription primers, the RNA having the poly (A) tail can be sequenced while the RNA with no poly (A) tail can be sequenced, the rRNA removal step is not required during the cell transcriptome sequencing, and the method is especially suitable for the sequencing on the transcriptome having the low initial RNA amount and the low cell number.

Description

technical field [0001] The invention relates to a transcriptome amplification and sequencing detection method, in particular to an amplification and sequencing technology capable of simultaneously covering ribonucleic acid species with and without polyadenylic acid tails in trace samples. Background technique [0002] With the development of life science research technology and the development of second-generation high-throughput sequencing technology, the research on organisms has become more and more detailed and in-depth. It is found that there is often heterogeneity between cells of the same species in the organism. The characteristics of these individuals may It is an important factor affecting and guiding the development of organisms, so the analysis at the single-cell level is becoming more and more important. In particular, the sequencing analysis of single-cell transcriptomes can directly reflect the overall expression differences between different cells and the reg...

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problem in the prior art that the RNA without poly(A) tail in the transcriptome cannot be sequenced, especially the RNA without poly(A) tail in the transcriptome of a small number of cells cannot be sequenced, the present invention provides A method that can reverse transcribe all kinds of RNA and amplify it to build a library, which uses "random primers" containing random sequences for reverse transcription, does not rely on poly(A) tails, so it can cover all ribonucleic acids type

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