A method for rapid cloning of physical quality regulation genes of elite winter sports athletes

A technology for physical quality and gene regulation, which is applied in the field of rapid cloning of excellent ice and snow athletes' physical quality regulation genes, and can solve problems such as limited length of amplified fragments, cumbersome RNA extraction process, and RNase contamination

Inactive Publication Date: 2013-09-11
HARBIN INST OF PHYSICAL EDUCATION
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AI Technical Summary

Problems solved by technology

The RNA extraction process is cumbersome, easily contaminated by RNase, and the length of the amplified fragment is limited; for larger genes, multiple amplifications are often required to obtain the full length

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0012] 1. Extraction of blood genomic DNA

[0013] (1) Take 20 μl of blood and put it in a 1.5ml centrifuge tube, add 500 μl of lysate.

[0014] (2) Cracking in an incubator at 55°C for 30 minutes.

[0015] (3) Add an equal volume of Tris-saturated phenol (phenol: chloroform: isoamyl alcohol 25:24:1), invert and mix well, then stand for extraction for 10 min, and centrifuge at 12000 g for 10 min at 4°C.

[0016] (4) Take the supernatant, add an equal volume of CI (chloroform: isoamyl alcohol 24:1), invert and mix well, then stand for extraction for 10 min, and centrifuge at 12000 g for 10 min at 4°C.

[0017] (5) Collect supernatants, add 2 times of absolute ethanol (stored at -30°C), invert and mix evenly, flocculent DNA can be observed. Precipitate in -30°C refrigerator for 30min. Centrifuge at 12000g for 10min at 4°C.

[0018] (6) Discard the supernatant, wash with 75% ethanol, and centrifuge at 7500 g for 5 min.

[0019] (7) Repeat the above operation once.

[0020] ...

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Abstract

The present invention proposes a method for rapid cloning of physical quality expression and regulation genes of elite winter sports athletes. Through extraction peripheral blood genomic DNA, and by using an improved overlap extension polymerase chain reaction (OE-PCR) technique to amplify physical quality related genes, products which are hard to get by using methods relying on restriction endonuclease digestion, can rapidly be obtained by using this technique. The method is time and effort saving, economical, accurate, efficient and simple for studying and exploring rapid cloning of physical quality expression and regulation genes of elite winter sports athletes, providing a most scientific method for the in-depth study of structures and functions of genes related to physical quality of winter sports athletes, and laying a solid theoretical and technical foundation for establishing a scientific system for early prediction of potential, evaluation and selection of winter sports athletes.

Description

technical field [0001] In recent years, with the rapid development of sports science and life science and the mutual intersection and penetration of theories, methods, technologies and means, it has strongly promoted the transformation and transformation of the early selection of ice and snow athletes to a scientific and systematic direction. develop. The present invention is a time-saving, labor-saving, economical, accurate, efficient and simple method for quickly cloning the expression and regulation of genes of excellent ice and snow athletes' physical fitness, which is suitable for the early selection of excellent ice and snow athletes at the gene level, and belongs to sports science and life sciences. Interdisciplinary disciplines and fields of science. Background technique [0002] Human genes are the most original data of life science research. Human genes can not only transmit genetic information to the next generation through replication, but also control protein s...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 朱志强赵玉华武大伟李向臣崔鹏关伟军
Owner HARBIN INST OF PHYSICAL EDUCATION
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