Mouse-tail DNA (deoxyribose nucleic acid) extraction kit applicable to the genotype of laboratory mouse and application thereof
A kit and genotype technology, applied in the field of accurate genotype identification of experimental mice, can solve the problems of increasing experimental costs, reagent costs, labor costs, overnight steps, unfavorable funds, etc., and achieve simplified routine extraction processes, reliable results, The effect of protecting integrity
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[0020] Example 1: Preparation of kit
[0021] A solution 40ml: Take 100μl 10N NaOH solution into a 50ml plastic container, then add 160μl 0.5M EDTA solution and 39.74ml ddH respectively 2 O, mix well.
[0022] B solution 40ml: Take 1.6ml 1M Tris solution and place it in a 50ml glass container, add 38.4ml ddH 2 O, mix well, pH 7.6.
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[0023] Example 2: Genotyping of Foxp3-GFP knock-in mice on C57B / 6 background (that is, knock green fluorescent protein into the mouse to link it with the Foxp3 gene, so that all cells expressing green fluorescence are Foxp3-positive cells)
[0024] The Foxp3-GFP knock-in adult mice paired with C57B / 6 background were gifted by Harvard University Medical School (purchased from Jackson Laboratory, USA), and placed in the Experimental Animal Center of the Third Affiliated Hospital of the Third Military Medical University, reared and bred in accordance with SPF animal standards , After weaning the pups, the Foxp3 gene was identified;
[0025] (1) Take the 0.2-0.5cm tissue from the tail tip of the mouse and place it in a 1.5ml EP tube;
[0026] (2) Add 180μl A solution;
[0027] (3) 100℃ 30min;
[0028] (4) Take out the EP tube and ice bath for 2 minutes;
[0029] (5) Add 180μl B solution and mix well;
[0030] (6) Store at 4°C for genotype identification:
[0031] PCR primer synthesis (Shangha...
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