Extraction buffer solution and extraction method for extracting DNA from bones or teeth

Abstract

The invention discloses a DNA extraction buffer solution which is formed by mixing a TES buffer solution, a sodium lauryl sarcosinate solution, a 1 mol / L DTT solution and a 10 mg / mL proteinase K solution in a volume ratio of (4-5):1:1:1. The invention also discloses a method for extracting DNA from bone or tooth samples in the presence of the DNA extraction buffer. The method comprises the following steps: cleaning impurities, pulverizing a sample at room temperature, adding the extraction buffer solution to cleave, adding an adsorption solution and adsorption beads for adsorption, rinsing adsorption beads, and adding an eluent for eluting. The invention also discloses a method of pretreating a sample during extraction of DNA from bones or teeth. The DNA extraction buffer solution and thecorresponding use method disclosed by the invention have the advantages thatthe efficiency of extracting DNA from bones and teeth is high, the extracted DNA is of good integrity and high purity, the operation is simple, rapid and safe, samples are free of crosscontamination, and the cost is low.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a DNA extraction buffer for extracting DNA from bones or teeth. The invention also relates to a method for extracting DNA from bones or teeth. Background technique [0002] Extracting DNA from bone or tooth samples has always been a technical challenge. Bones and teeth are relatively special biological specimens. Taking bone tissue as an example, there is a large amount of calcified matrix outside the bone cells, which provides good mechanical strength for the bone, but this also leads to insufficient exposure of the bone cells during the DNA extraction process, and cannot be in good contact with the lysate. Makes DNA extraction ineffective. Similarly, when extracting DNA from dense dental tissue, there is also the problem of low extraction efficiency. [0003] In order to improve the extraction efficiency of DNA in bones or teeth, there are two main methods generally adopted in th...

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that it takes a long time and will cause DNA loss, which is very unfavorable for precious samples with small sample sizes

This method is cumbersome and laborious to operate, and liquid nitrogen is required when using it, which may cause the risk of frostbite to the operator

Alternatively, other existing mechanical crushing methods include using an electric drill to drill bone powder, or crushing teeth with a smashing method, all of which have certain risks and limitations.

For example, the refrigerated grinder of the old brand product 6770 of SPEX company in the United States is to immerse the sample bottle in liquid nitrogen, and use electromagnetic as the power to drive the steel impactor to crush and grind. The maximum size of the grinder is only 25mm. The applicability is poor, the price of the machine and grinding consumables is very expensive, and the grassroots unit cannot afford it, and the sample bottle needs to be cleaned and reused repeatedly, and there is a problem of easy cross-contamination between samples

[0006] In addition, in recent years, Life Technologies has introduced a DNA extraction kit based on the BTA method. This kit has improved the extraction speed, but there are still deficiencies in the extraction quality and the price is expensive.

Images