Kit for extracting DNA for sampling in minute quantities without wound, and method for extracting DNA
A kit and micro-quantity technology, applied in the field of biological DNA detection, can solve the problems of difficult and sensitive human sampling, and achieve the effects of reducing costs, saving resources, and improving quality and quantity
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Embodiment 1
[0045] Embodiment 1, the configuration of each component of kit
[0046] Solution composition:
[0047] (1) Lysis solution: GuSCN (guanidine isothiocyanate), EDTA, Tris-HCl, Triton X-100;
[0048] (2) Washing solution A: GUSCN (guanidine isothiocyanate), Tris-HCl;
[0049] (3) Washing solution B: 70% ethanol;
[0050] (4) washing liquid C: acetone;
[0051] (5) Eluent: Tris-Hcl, EDTA (sterilized).
Embodiment 2
[0052] Embodiment 2, human hair and saliva DNA extraction
[0053] First perform non-invasive microsampling:
[0054] (1) Hair: Pull out the hair of the experimental subject, take 1, 3, and 5 hairs respectively, cut about 0.5-1 cm from the base of the hair root with clean and dry scissors, and place them in a 1.5 mL eppendorf tube;
[0055] (2) Saliva: Use a 1.5 mL eppendorf tube to directly collect about 0.3 mL of the subject's saliva.
[0056] The specific extraction steps are as follows:
[0057] (1) Add 1mL of lysate respectively, and bathe in water at 56°C for 1h;
[0058] (2). Add 20 μl of silica gel adsorbent, mix well, and fully transpose;
[0059] (3) After sufficient centrifugation, discard the supernatant;
[0060] (4) Resuspend in 1mL washing solution A, centrifuge thoroughly, and discard the supernatant. Repeat this step once;
[0061] (5) Resuspend twice in 1mL washing solution B and centrifuge fully;
[0062](6) Resuspend once in 1mL washing solution C an...
Embodiment 3
[0066] Embodiment 3, the inspection of DNA extraction product
[0067] The extraction product obtained in Example 2 is measured at 260nm by a UV spectrophotometer, and it can be calculated that the DNA extraction amount of 1, 3, and 5 hairs is about 100ng, 300ng, and 500ng respectively, and the DNA extraction amount of 0.3mL saliva is about 100ng. It is 2.0ug, converted into the number of experiments are 10, 30, 50, 200 times. The gel electrophoresis profile of the extract is shown in Figure 4.
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