Process for extracting cytoplasm DNA
An extraction method and cytoplasmic technology, applied in the field of DNA extraction, can solve the problems of unstable cytoplasmic DNA quality, time-consuming and experimental materials, cumbersome operation process, etc., and achieve the effects of low extraction cost, high success rate and improved extraction efficiency.
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Embodiment 1
[0029] (1) Preparation of cytoplasmic DNA
[0030] Grind the human lung cell tissue, filter the tissue cells with a metal strainer to obtain single cells, use TBS liquid (the configuration method of TBS liquid: get NaGl 8g, KCl 0.2g, Tris 3g, add deionized double distilled water 800ml to dissolve, use 2mol / L HCl to adjust the pH value to 7.4, distilled water to 1000ml.) Wash twice, centrifuge at 2000r / min, discard the supernatant, and then use STE solution (the configuration method of STE solution: take 5.84g NaCl, 0.5mol / L Tris-HCl 20ml, 0.5mol / LEDTANa 2 2H 2 Dissolve 2ml of O in 800ml deionized double-distilled water, adjust the pH value to 7.4 with 2mol / L HCl, and adjust the volume to 1000ml with double-distilled water. ) was washed once, the supernatant was discarded by centrifugation, and the number of cells in the pellet was 10 7 . Add solution A at 4°C to the cell pellet (the preparation method of solution A: take 1g of glucose, 5ml of 0.5mmol / L Tris-HCl, 0.5mol / L ...
Embodiment 2
[0035] Extract 1-3ml of heparinized anticoagulated peripheral venous blood from the human body, centrifuge at low speed for 5min, take buffy layer, wash twice with TBS solution (the configuration method of TBS solution is the same as in Example 1), centrifuge at 2000r / min, discard the supernatant, and use The STE solution (the configuration method of the STE solution is the same as in Example 1) was washed once, and the supernatant was discarded by centrifugation, and the number of cells in the sediment was 10 7. Add 140 μl of solution A at 0°C (the configuration method of solution A is the same as in Example 1) to the cell pellet, blow and mix in an ice bath, and then add 290 μl of solution B at 15°C (the configuration method of solution B is the same as in Example 1), Gently invert and mix well, cleavage and denature in ice bath for 5 minutes, then add 215 μl of 0°C solution C (the preparation method of C solution is the same as in Example 1), mix gently, and refold in ice b...
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