Method for treating and crushing plant single cell sequencing sample

Abstract

The invention discloses a method for treating and crushing a plant single cell sequencing sample. The method comprises the following steps: carrying out physical crushing treatment on a plant sample; enzymatic hydrolysate is added into the plant sample to obtain an enzymolysis mixture, the enzymolysis mixture is subjected to enzymolysis under the condition that the pH value is 6.0 and the temperature is controlled to be 25-35 DEG C, and the enzymatic hydrolysate is prepared by mixing cellulase, pectinase, calcium chloride, mannitol and macerozyme R-10 according to the mol ratio of 30: 1: 50: 600: 1; and filtering the enzymolysis mixture, separating a single cell or a plurality of groups of cells, and carrying out an on-machine pre-program for sequencing a sample. According to the method, the cells are promoted to be dissolved into water from a biological sample in the form of physical crushing and enzymolysis, the yield and purity of the cells are improved, the preparation method of the plant single cell transcriptome sequencing sample is established and optimized by adopting multiple methods, and the obtained plant cells are high in purity and activity and convenient to detect.

Description

technical field [0001] The invention belongs to the technical field of plant single cell detection, and in particular relates to a method for processing and crushing plant single cell sequencing samples. Background technique [0002] Biological resources are important resources for studying the origin, variation and development direction of organisms. Whether it is a single-celled organism or a single cell isolated from a biological tissue, it has important research value. Typical single-cell biological resources include bacteria, fungi, archaea, etc., and the number and types of these microbial cells are various. Functional microorganisms of different species are widely used in food fermentation, organic matter degradation and other fields. Many functional microbial single cells exist in animal and plant tissues, mixed with other impurities such as protein and fat, and it is difficult to separate impurities. [0003] Living matter such as animals and plants are advanced l...

AI Technical Summary

Problems solved by technology

However, multiple cell proteins in animal and plant tissues stick together, and the viability of single cells is far inferior to that of microorganisms, and it is also difficult to separate impurities

[0004] As a cutting-edge technology, single-cell sequencing is widely used in animal and human cells, but it is rarely used in the field of plants

To sum up, the problem existing in the prior art is that the size of most single cells is mostly between a few microns and tens of microns, the diameter is small, and it is difficult to separate biological cells from impurities, resulting in low cell purity. Sequencing sample processing fragmentation method

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