Lahiri D K, Schnabel B
Department of Psychiatry, Indiana University School of Medicine, Indianapolis 46202-4887.
Biochem Genet. 1993 Aug;31(7-8):321-8. doi: 10.1007/BF02401826.
The isolation of DNA from whole blood by a modified rapid method (RM) was tested using various detergents and buffer conditions. Extraction of DNA with either NP-40 or Triton X-100 gave a high yield of undegraded DNA in less than an hour. The concentration of magnesium ion in the buffers was critical to obtaining intact, high molecular weight (HMW) DNA. Greater than 10 mM MgCl2 led to degradation. Addition of EDTA to the buffer inhibits this degradation. Preparation of DNA from blood stored at room temperature or incubated at 37 degrees C for 24 hr resulted in the same amount and quality of DNA as from samples frozen at -70 degrees C. DNA from blood samples that had undergone more than four freeze-thaw cycles was found to be partially degraded. The modified RM can be applied to extract DNA from as little as 10 microliters of blood (340 ng of DNA) and from dried blood samples. DNA samples remained intact and undegraded for longer times when DNA was dissolved in higher concentrations of EDTA.
使用各种去污剂和缓冲液条件，对通过改良快速方法（RM）从全血中分离DNA进行了测试。用NP - 40或Triton X - 100提取DNA，在不到一小时的时间内可获得高产率的未降解DNA。缓冲液中镁离子的浓度对于获得完整的高分子量（HMW）DNA至关重要。大于10 mM的MgCl₂会导致降解。向缓冲液中添加EDTA可抑制这种降解。从室温保存或在37℃孵育24小时的血液中制备DNA，其DNA的量和质量与从-70℃冷冻的样品相同。发现经过超过四个冻融循环的血液样本中的DNA部分降解。改良的RM可应用于从低至10微升的血液（340 ng DNA）和干血样本中提取DNA。当DNA溶解在更高浓度的EDTA中时，DNA样本可在更长时间内保持完整且未降解。
