Long-term storage and RNA extraction method of whole blood sample

Abstract

The invention discloses a long-term storage and RNA (ribonucleic acid) extraction method of a whole blood sample, which comprises the steps of immediately mixing acquired fresh whole blood with 5-15 times of volume of a TRIzol reagent, storing a whole blood-TRIzol mixed system in a refrigerator at -80 DEG C after homogenization, after storage for a period of time, extracting total RNA from the stored whole blood-TRIzol mixed system. The method achieves direct long-term storage of the human whole blood after the homogenization with no need for plasma separation or red blood cell lysis, and whole blood subpackage, and high-quality total RNA with very high purity and integrity is extracted.

Description

technical field [0001] The invention relates to a blood sample preservation and RNA extraction method, in particular to a method for long-term preservation of human whole blood samples used for RNA extraction and RNA extraction. Background technique [0002] With the completion of the Human Genome Project in the early 21st century, life science has entered the post-genome era, and new omics studies such as transcriptomics, proteomics, and metabolomics have emerged as the times require. In the central dogma, RNA is the bridge connecting DNA and protein, reflecting the dynamic changes in the functional state of cells. Transcriptomics studies gene expression at the RNA level and plays an important role in exploring the biological mechanisms of disease occurrence and development. In recent years, it has been more and more applied in clinical medicine, epidemiology, molecular biology and other fields. The extraction of RNA is the basis of experiments such as RNA-seq, microarray...

AI Technical Summary

Problems solved by technology

Very heavy workload for studies collecting large numbers of biological samples in a short period of time

[0007] 3. Due to the influence of RNA's own structure and RNase and other factors, the existing technology requires RNA to be extracted as soon as possible after blood collection, which increases the storage time of whole blood and increases the risk of RNA degradation

[0008] 4. The purity of whole blood RNA extracted by existing technology is low

The extraction process is mostly carried out in 1.5mL centrifuge tubes without RNase. The system is small, and carbohydrates (sugars), salts, or organic solvents may be left in the washing process, making OD260 / 230 (an indicator indicating RNA purity) ) lower

For a large amount of whole blood, it needs to be divided into several tubes for extraction, which increases the operation steps and time, and easily introduces risk factors that affect the quality of RNA

[0009] 5. In the drying process after RNA washing, the natural drying time is longer, which increases the possibility of RNA exposure to RNase in the environment, resulting in an increased risk of degradation

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