Method for extracting a large number of high-purity deoxyribonucleic acid (DNA) from chicken blood

Abstract

The invention discloses a method for extracting a large number of high-purity deoxyribonucleic acid (DNA) from chicken blood. The method comprises absorbing an unfreezing chicken blood sample preserved through DNA preserving fluid into a centrifugal tube containing a reagent X; performing repeated dangling through a liquid-moving gun until the original blood sample becomes clear liquid; performing centrifugation and removing supernate; adding a reagent Y, slightly performing suspension on centrifuged sediment obtained by the last step; adding ribonuclease (Rnase) A, evenly mixing and then performing warm bath at the temperature of 37 DEG C for 30 minutes; adding 5mol / L of sodium perchlorate, performing manual oscillation to achieve mixing for 5 minutes; performing water bath, manually shaking the mixture for one time every 5 minutes; adding precooled chloroform, mixing for 5 minutes, centrifuging and absorbing supernate into another centrifugal tube; adding isopyknic isopropyl alcohol, and slowly shaking to obtain flocculent DNA; washing the flocculent DNA through precooled 75% of ethanol for two times, airing, performing dissolution through appropriate TE (mixture of tris and ethylene diamine tetraacetic acid ), storing for one week at the temperature of 4 DEG C and then storing for a long term at the temperature of -20 DEG C. By means of the method, the time for extracting the DNA is short, the extracted DNA sample has high purity, large concentration and good quality, and the high-purity deoxyribonucleic acid (DNA) can completely meet requirements of downstream molecular biology experiments.

Description

technical field [0001] The invention relates, in particular, to a method for extracting a large amount of high-purity DNA from chicken blood. Background technique [0002] Extracting high-quality, high-concentration and high-purity genomic DNA is an important technical step in carrying out molecular biology experiments. Extracting genomic DNA from animal blood has the advantages of not killing animals, economical and practical. Conventional blood DNA extraction methods mostly use phenol-chloroform extraction, which has the disadvantages of time-consuming, highly toxic experimental reagents, and complicated operations. The extraction cost of the kit is relatively high, and the purity and concentration of the extracted DNA are low, which is only suitable for the needs of recent experiments. The small amount of extracted DNA is not conducive to repeated use; and due to the small amount of extraction, the extracted DNA is easy to degrade and difficult Long-term storage and util...

AI Technical Summary

Problems solved by technology

Conventional blood DNA extraction methods mostly use phenol-chloroform extraction, which has the disadvantages of time-consuming, highly toxic experimental reagents, and complicated operations.

The extraction cost of the kit is relatively high, and the purity and concentration of the extracted DNA are low, which is only suitable for the needs of recent experiments. The small amount of extracted DNA is not conducive to repeated use; and due to the small amount of extraction, the extracted DNA is easy to degrade and difficult Long-term storage and utilization, the downstream PCR amplification reaction cannot be performed if the time is longer

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