The invention belongs to the field of
transcriptome analysis and relates to a quick single-
cell RNA (ribonucleic acid) reverse transcription and
library construction method. According to the method, 20-500ng high-quality full-length double-chain cDNA (
complementary deoxyribonucleic acid) is amplified by taking 1-2000 cells or 10pg-20ng extracted
eukaryote total RNA as an initial, and a high-quality
cDNA library meeting downstream analysis requirements is obtained. The method effectively avoids 3' preference and
genome DNA contamination in a cDNA synthesis process; an expression quantitation molecule
label can assist in
gene expression quantity calculation; and at the same time, the expression quantitation molecule
label can also keep chain source information during complete amplification of
RNA sequence information. The method can achieve a reverse transcription and amplification
library construction success rate of above 95%; the
cDNA library can be connected with an Illumina main
stream sequencing platform; lower computer data (5M Reads) can detect
gene expression of above 90%; the
gene expression consistency exceeds 90%; amplification has no obvious bias; and a required sample input amount is smaller.