dna提取原理和方法-文庫吧
2025-04-20 10:02 本頁面
【正文】 ix well and incubate at 55186。C overnight or until tissue is dissolved。 More Proteinase K may be added if digestion is not plete after 24 hr。 ? Add ml Phenol/Chloroform and vortex at top speed for 15 min at RT。 ? Spin at 13,000 rpm for 30 min at RT。 ? Transfer the top aqueous phase to a fresh microfuge tube。 Add ml Chloroform and vortex at top speed for 5 min at RT。 Spin at 13,000 rpm for 5 min at RT。 ? Transfer the top aqueous phase to a fresh microfuge tube, add 50 ml 3 M NaOAc (pH=) and ml 100% ethanol, invert several times。 A NaOAc solution with pH lower than will cause the EDTA to precipitate。 Spin at 13,000 rpm for 5 min at RT。 If there is no pellet, place samples in 80186。C for 10 min and spin at 13,000 rpm for 25 min at 4186。C。 ? Wash pellet once with 70% ethanol, air dry。 ? Resuspend DNA in ddH2O. ? Use ~100200 ml ddH2O to get final concentration as 100ng/ml for PCR. Phenolchloroform extraction of mouse tail DNA ： 十二烷基硫酸鈉 ： 緩沖液 2. Add ml tail solution and 25 ml Proteinase K, mix well and incubate at 55186。C overnight or until tissue is dissolved。 作用：細(xì)胞裂解時 PH值也能穩(wěn)定 作用： a. 溶解細(xì)胞膜、核膜上脂質(zhì)成分 作用：是二價金屬螯合劑，抑制核酸酶；降低細(xì)胞膜的穩(wěn)定性。 Tail solution: 3. More Proteinase K may be added if digestion is not plete after 24 hr。 Proteinase K :
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