Single cell transcript isomer sequencing analysis method and kit

Abstract

The invention discloses a single cell transcript isomer sequencing analysis method and a kit. By adding tag sequences to the two tail ends of a single mRNA, after trapping and enriching are performed,the whole transcriptome is sequenced, and by accurately judging the head and tail structures of different transcript isomers, full-length transcripts can be judged. Existing technical problems can besolved by the new research strategy. The single cell transcript isomer sequencing analysis method and the kit have the following beneficial effects that 1, the sensitivity of the detection of the full-length transcripts is improved, specifically, due to the targeted detection of the head and tail areas of the transcripts, signals are mostly concentrated at the end edges of the transcripts, so that the detection of the full-length transcripts is higher; 2, simple operation is achieved, and the cost low, specifically, the method is simple to operate, widely applicable and the cost is low; and 3, the accuracy of the detection of the transcripts is improved, specifically, higher accuracy can be achieved by combining the tag sequences of the tail end sequences and the optimization of a data analysis algorithm.

Description

Technical field: [0001] The invention belongs to the field of biomedicine, and in particular relates to an analysis method and a kit for sequencing single-cell transcript isoforms. Background technique: [0002] The variability of gene transcription isoforms leads to the diversity of gene functions, and the different selection of mRNA at the transcription initiation site and transcription termination site can lead to differences in mRNA stability, positioning and translation efficiency, and even produce truncations with different functions type protein. Studies have confirmed that there is significant heterogeneity in function and molecular characteristics among single cells that make up human organ tissues. Existing single-cell sequencing methods can distinguish gene expression differences between different cells, but most of these methods are based on the 5' of mRNA Or capture sequencing at the 3' end instead of detecting full-length transcripts. The two currently report...

AI Technical Summary

Problems solved by technology

The two currently reported single-cell full-length transcript sequencing methods have problems such as low sensitivity and high cost. For example, smart2-seq can detect the full-length single-cell transcriptome, but most of the detection signals are distributed in the middle part of the gene. However, there is very little distribution at the end of the transcript, resulting in low sensitivity and accuracy for full-length transcript analysis

Pacbio long-read and long-term third-generation sequencing can perform full-length sequencing of mRNA transcripts, but there are problems such as high cost and relatively high error rate

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