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A kit and extraction method applicable to high-throughput extraction of genomic DNA from animal tissues

An animal tissue and high-throughput technology, which is applied in the field of molecular biology, can solve the problems of unsatisfactory genomic DNA purity and high requirements of the lysis effect, and achieve scientific and reasonable composition and proportioning, high requirements, and improved efficiency. Effect

Active Publication Date: 2021-10-12
SHANDONG HONGJITANG PHARMA GRP JINAN E JIAO PROD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the lysing effect of the DNA extraction kits currently on sale on animal skins and the purity of the extracted genomic DNA cannot well meet the requirements of the subsequent detection and analysis steps, especially the automated high-throughput DNA extraction has higher requirements for the extraction kits

Method used

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  • A kit and extraction method applicable to high-throughput extraction of genomic DNA from animal tissues
  • A kit and extraction method applicable to high-throughput extraction of genomic DNA from animal tissues
  • A kit and extraction method applicable to high-throughput extraction of genomic DNA from animal tissues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A kit applicable to high-throughput extraction of genomic DNA from animal tissues, including lysate, binding solution, washing solution I, washing solution II, eluent, proteinase K and magnetic beads;

[0040] The lysate is made of the following components: 3M guanidine isothiocyanate, 10mM EDTA, 2% (mass percentage) SDS, 25mM NaCl, 0.5% (volume percentage) Triton-X100, 2% (volume percentage) Tween-20 , the pH of the lysate was adjusted to 7.8 with dilute hydrochloric acid;

[0041] The binding liquid is made of the following components: 3M guanidine isothiocyanate, 1.0M NaCl, 1% (mass percentage) Glucose, 35% (volume percentage) ethanol, and the pH of the binding liquid is adjusted to 6.5 with dilute hydrochloric acid;

[0042] The washing solution is composed of washing solution I and washing solution II; the washing solution I is made of the following components: 4M urea, 0.5M NaCl, 50% (volume percentage) ethanol, and the pH of the washing solution I is 7.5; The wa...

Embodiment 2

[0062] A kit applicable to high-throughput extraction of genomic DNA from animal tissues, including lysate, binding solution, washing solution I, washing solution II, eluent, proteinase K and magnetic beads;

[0063] The lysate is made of the following components: 6M guanidine isothiocyanate, 50mM EDTA, mass ratio 2%SDS, 25mMNaCl, volume ratio 2%Triton-X100, volume ratio 5%Tween-20, the pH of the lysate is diluted with dilute Hydrochloric acid is adjusted to 7.8;

[0064] The binding solution is made of the following components: 6M guanidine isothiocyanate, 2M NaCl, 2% Glucose by mass ratio, and 50% ethanol by volume, and the pH of the binding solution is adjusted to 6.5 with dilute hydrochloric acid;

[0065] The washing solution is composed of washing solution I and washing solution II; the washing solution I is made of the following components: 8M urea, 0.5M NaCl, 50% ethanol by volume, and the pH of the washing solution I is 7.5; the washing solution II is made of the fol...

Embodiment 3

[0083] Adopting the kit of the present invention, extracting with a high-throughput DNA automatic nucleic acid extractor includes the following steps:

[0084] (1) prepare each solution by above-mentioned amount, after autoclave sterilization, standby;

[0085] (2) Take 192 animal tissue samples (each 1.5*0.4cm2), (the samples are non-soaked samples that have been dried for a long time), put them in two 96-well square well plates, add 600uL lysate, 400ug respectively Proteinase K, shake and incubate at 900rpm at 56°C for 4h, and extract DNA according to the following steps;

[0086] (3) Centrifuge 2 plates of 96-well square well plates in which the sample in step (2) is completely dissolved at 12000 rpm for 10 min;

[0087] (4) Put the centrifuged 2 plates of 96-well square-well plates in the DNA nucleic acid extractor, pour the prepared binding solution + magnetic beads (60:1), washing solution Ⅰ, and washing solution Ⅱ into the solution tank, Put the 96 round hole plate, m...

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Abstract

The invention relates to the technical field of molecular biology, in particular to a kit and an extraction method applicable to high-throughput extraction of genomic DNA from animal tissues. The kit, which can be applied to high-throughput extraction of genomic DNA from animal tissues, includes lysate, binding solution, washing solution I, washing solution II, eluent, proteinase K and magnetic beads. By lysing and destroying cells to remove proteins, high-purity DNA is obtained. Compared with commercially available kits, the present invention has fast lysis speed, high extraction purity, and large extraction volume. The reagents used are safe, stable and efficient, and are especially suitable for high-throughput automated production. The extraction efficiency of genomic DNA is greatly improved, the error is reduced, and it is more accurate.

Description

Technical field: [0001] The invention relates to the technical field of molecular biology, in particular to a kit and an extraction method applicable to high-throughput extraction of genomic DNA from animal tissues. Background technique: [0002] Donkey skin is the dry skin or fresh skin of equine animal donkey (Equus asinus L.), which is the raw material for preparing traditional Chinese medicine donkey-hide gelatin. Donkey-hide gelatin is the treasure of health care in China and has a long history. The Han "Shen Nong's Materia Medica" is listed as the top grade, and the Ming "Compendium of Materia Medica" is listed as the best nourishing product, the product of flesh and blood, the most nourishing, and has been used as a beauty product for women in all dynasties. Donkey-hide gelatin has the functions of nourishing qi and blood, enhancing physical fitness, strengthening the heart and lungs, strengthening muscles and bones, and beautifying the skin. In recent years, with t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 李凯刘子夜郝大魁
Owner SHANDONG HONGJITANG PHARMA GRP JINAN E JIAO PROD CO LTD
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