Multi-omics method for single cell transcriptome and translation group combined sequencing

Abstract

The invention provides a method for simultaneously carrying out transcriptome sequencing and transscriptome sequencing, which is characterized by comprising the following steps: carrying out first-step lysis treatment on cells and a lysis buffer solution to obtain a first lysis solution; after uniformly mixing and centrifuging, taking a part of the first lysis solution in the supernate as a transcriptome template, and taking the rest of the supernate and the lysis buffer solution to carry out second lysis treatment so as to obtain a second lysis solution; centrifuging the second lysate, and collecting supernate as a translation group template; obtaining a transcriptome cDNA library by using the transcriptome template, and sequencing the transcriptome cDNA library; and extracting a ribosome protected RNA fragment from the translation group template, and carrying out library establishment and sequencing on the ribosome protected RNA fragment. According to the method, high-quality transcriptome and translation group data in a whole genome range can be obtained at the same time, and the method is particularly suitable for single cells, so that possibility is provided for knowing collaborative regulation of single cell transcription and translation levels.

Description

technical field [0001] The present invention relates to the field of biology. In particular, the present invention relates to methods for performing transcriptome sequencing and translationome sequencing simultaneously. Background technique [0002] In recent years, with the development of next-generation sequencing technology, the emergence of many high-throughput sequencing technologies is helping people gain an in-depth understanding of various processes in biology. For research related to gene expression regulation, a large number of omics technologies based on the measurement of DNA and mRNA levels have emerged. Among them, the single-cell transcriptome method was first established in 2009, has undergone a series of optimizations, and has been widely used in various biological research processes today. However, single-cell transcriptome sequencing cannot accurately reflect the abundance of proteins encoded by each gene in the cell, because the translation efficiency a...

AI Technical Summary

Problems solved by technology

However, due to technical bottlenecks, the existing single-cell multi-omics technology mainly focuses on the combination of DNA, mRNA, DNA and chromatin modification, but there are few studies on protein translation or protein level.

Although some research groups have tried to combine the transcriptome and proteome based on antibody-labeled protein antigens, they can only detect dozens of proteins with high abundance, and cannot link translation and transcription of proteins at the genome-wide level. to study together

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