Kit capable of being applied to high-throughput extraction of genome DNA from animal tissue and extraction method thereof
An animal tissue and high-throughput technology, applied in the field of molecular biology, can solve the problems of unsatisfactory lysis effect, genomic DNA purity and high requirements, and achieve scientific and reasonable component composition and ratio, high requirements, and short time-consuming Effect
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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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