Storing and RNA extracting method of human blood sample

Abstract

The invention belongs to the technical field of biology, and relates to a storing and RNA extracting method of a human blood sample. The method comprises the following steps: collecting blood througha negative pressure blood sampling tube which is filled with an anticoagulant and a Trizo1 agent; storing under room temperature; extracting RNA, wherein the ratio of the anticoagulant to the Trizo1 agent to negative pressure in the blood sampling tube is 0.8-1.2 mL to 5.5-8.0 mL to 2.8-4.0 mL; the blood is immediately protected once after being sampled; the blood is stored for 30 days under roomtemperature; the RIN in the extracted RNA is beyond 8.0, the completeness of RNA is high, and the purity is high; the RNA extracting method is simple and can be finished within 15min; the extracting rate is high; the blood sampling volume is reduced. Therefore, the blood is sampled through the negative pressure blood sampling tube and is stored under room temperature, so that the extracting methodis simplified, and cold chain transportation is avoided; the cost is decreased; the method is convenient to popularize.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for preserving human blood samples and extracting RNA. Background technique [0002] With the rapid development of molecular biology, there are more and more researches on nucleic acids. Human blood is one of the most easily obtained clinical samples and is widely used in medical research, especially in clinical diagnosis. The extraction of high-quality total RNA from blood samples is the basis for downstream sequencing, qPCR, chip detection and other technologies. However, there are technical difficulties in extracting high-quality total RNA, and RNA preservation is a major bottleneck in the study of gene expression groups. First of all, RNA is very fragile and extremely difficult to preserve. Due to the widespread presence of RNases in nature, including RNases in tissues and pathogens, RNA begins to degrade within a few minutes at room temperature, and completely degrade...

AI Technical Summary

Problems solved by technology

This method first processes blood samples, adds Trizol reagent to preserve whole blood cells, and does not need to add Trizol in the subsequent RNA extraction process, which simplifies the steps of Trizol method for RNA extraction, but increases the complexity of blood sample processing and requires blood collection. There are RNAfree consoles, centrifuges, pipettes and professional technicians on site, making it impossible to use in primary hospitals or collect a single sample

In addition, the quality of RNA extracted by this method is unstable, and the RIN value is only above 7.7, while RNA sequencing requires an RIN value of at least 8.0; the pretreated blood still needs to be stored at -80°C, and the operation is complicated and requires Cold chain transportation, high cost

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