Lysis solution and method for extracting ancient biological bone DNA

A technology of lysate and bone, which is applied in the field of biological DNA extraction, can solve the problems of high impurities in samples, high toxicity of human body, incomplete tissue lysis, etc., and achieve the effect of improving the effect of amplification and the degree of lysis

Active Publication Date: 2016-03-23
CENT SOUTH UNIV
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Problems solved by technology

However, some existing old methods have shortcomings such as incomplete tissue lysis, more DNA loss, and too many impurities in the obtained samples, the phenol / chloroform method is highly toxic to the human body, and even the decalcification method also has disadvantages such as taking too long

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  • Lysis solution and method for extracting ancient biological bone DNA
  • Lysis solution and method for extracting ancient biological bone DNA
  • Lysis solution and method for extracting ancient biological bone DNA

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Embodiment Construction

[0033] The following in conjunction with specific embodiments is intended to further illustrate the present invention, rather than limit the present invention.

[0034] The new lysate formula provided by the present invention: pH8.5 Tris-Hcl, 10mM; EDTA, 15mM; SDS, 1.5%;

[0035] NaCl, 150 mM; Proteinase K, 0.5 mg / ml.

[0036] Other reagents and consumables: isopropanol, DNA spin column, protein removal solution, DNA washing solution, absolute ethanol, etc.

[0037] The steps that the present invention implements:

[0038] (Before operation, add 3 times the volume of absolute ethanol to the rinse solution in advance)

[0039] 1. Cut out a small piece of unearthed ancient human skeleton, crush it with a grinder, weigh 0.3g and put it into a 1.5ml EP tube. The remaining powder can be collected and stored at -20°C;

[0040] 2. Add 300 μl of lysate and incubate at 55°C for 12–16 hours;

[0041] 3. Centrifuge at 12000rpm for 10min, take 250μl of the supernatant into a new EP tu...

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Abstract

The invention discloses a lysis solution and method for extracting ancient biological bone DNA. The lysis solution is prepared from 10 mM of Tris-HCl with pH of 8.5, 10-15 mM of EDTA, 1.0-1.5 wt% of SDS, 150 mM of NaCl and 0.3-0.5 mg/ml of protease K. By means of improving the components of the SDS, the EDTA and the protease K respectively, the lysis degree of the lysis solution on bone tissue can be obviously improved. Meanwhile, when the lysis solution is used, by means of protein solution removing, DNA wash liquid treatment and a DNA centrifugal column method, impurity composition in DNA samples can be obviously reduced, the improvement of PCR amplification effect is facilitated, and therefore high-purity ancient biological bone DNA can be obtained conveniently.

Description

technical field [0001] The invention belongs to the technical field of biological DNA extraction, and in particular relates to a lysate and a method for extracting ancient biological bone DNA. Background technique [0002] Extracting DNA from ancient biological bones is of great significance for exploring the life mysteries of ancient organisms and the development of human civilization. Bone is a special biological detection material, because it has a hard protective tissue on the outside, so it degrades slowly compared to other tissues and organs of the organism. Therefore, for highly decomposed samples, bone is one of the effective detection materials for DNA detection. Due to the action of nucleases and microorganisms in the surrounding environment, the DNA strands in ancient biological bones will be broken, missing or even degraded, making it difficult to obtain sufficient DNA samples. Therefore, there is still a need to find a more effective DNA extraction method to m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 李志远马文波邓思浩李彩月陈旦刘亮
Owner CENT SOUTH UNIV
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