Sample treatment device used before unicellular sequencing, micro-fluidic chip and application

Abstract

The embodiment of the invention provides a sample treatment device used before unicellular sequencing. The device comprises a first main flow channel, a second main flow channel and at least one sample treatment unit; the first main flow channel is provided with at least one branch flow channel, and a sample input port and a sample output port are formed in the two ends of the first main flow channel respectively; one sample treatment unit is arranged on each branch flow channel, and comprises a cell capturing cavity, a nucleic acid releasing cavity, a neutralization reaction cavity and a liquid droplet generating cavity, and the cell capturing cavity, the nucleic acid releasing cavity, the neutralization reaction cavity and the liquid droplet generating cavity are communicated with one another in sequence; a control valve is arranged between every two adjacent cavities; the second main flow channel is communicated with the cell capturing cavity in each sample treatment unit and provided with a cell input port. By means of the device, unicellular whole-genome amplification can be efficiently achieved, and the uniformity and accuracy of a unicellular whole-genome amplification reaction are greatly improved. The invention further provides a micro-fluidic chip and application.

Description

technical field [0001] The invention relates to the field of microfluidic technology, in particular to a sample processing device before single cell sequencing, a microfluidic chip and its application. Background technique [0002] Single-cell whole-genome sequencing (single-cell sequencing for short) technology is a new technology for amplifying and sequencing the whole genome at the single-cell level; its principle is to amplify the whole-genome DNA of an isolated single cell to obtain high coverage Following the rate of complete genomes, exon-capture high-throughput sequencing is used to reveal differential relationships among cell populations. In recent years, the most common and simplest method for amplifying the genomic DNA of a single cell is the multiple displacement amplification (MDA) technique, which uses random primers and isothermal amplification to obtain a large amount of high-fidelity DNA fragments, but there are problems such as amplification bias and non-s...

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Problems solved by technology

However, in this process, the genetic material is easily degraded and lost or polluted by external sources, which greatly increases the difficulty of the operation, which brings great challenges for researchers to further understand the heterogeneity at the single-cell level and its biomedical significance. great challenge

[0004] If the original amount of whole genome amplification PCR that has been reported so far is reduced to a single cell, accurate amplification cannot be obtained, so there are obvious technical barriers

At the same time, in the existing method technology, the coverage rate of most single-cell sequencing reaches about 40%-70% of the genome

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