A fast procedure for yeast DNA purification.
A fast procedure for yeast DNA purification.
复制标题
酵母 DNA 纯化的快速程序。
DOI：
10.1093/nar/19.19.5443
复制
发表时间：
1991
影响因子：
14.9
通讯作者：
A. C. Adam
中科院分区：
文献类型：
--
作者：
Julio Polaina;A. C. Adam
据报道，用3% SDS处理细胞悬液以引起细胞裂解，是一种有效的分离枸橼酸革兰氏阳性菌DNA的简单方法（1）。我们已经应用该方法分离酵母DNA，克服了标准方案中包括的麻烦的原生质体生成（2）。用于DNA纯化的方案是野生型酵母在液体YPD培养基中过夜。在1.5ml微量离心管中离心收集的1 ml等分试样的细胞，重悬于100 μ L TE缓冲液（10 mM）中，并在室温下保持15分钟，偶尔搅拌。通过向每个管中加入0.5 ml TE稀释破碎的细胞;这样做是为了在下一步骤中用苯酚处理后进行相分离。通过用1体积用TE平衡的液体苯酚处理来进行脱蛋白。用乙醚处理水相中的核酸以消除残留的苯酚，并通过异丙醇回收，通过处理（DNA酶，tg/4 t1，tg
The treatment of a cell suspension with 3 % SDS, to cause cell lysis, has been reported effective as a simple method for the isolation of DNA from recalcitrant gram-positive bacteria (1). We have applied this procedure to isolate yeast DNA, overcoming the troublesome generation of protoplasts included in standard protocols (2). and for yeast cultivation The protocol used for DNA purification was Saccharomyces wild type overnight in liquid YPD medium. The cells from 1 ml aliquots of the collected centrifugation in 1.5 ml microfuge tubes, resuspended in 100 of TE buffer (10 mM 1 containing and maintained at room temperature for 15 minutes with occasional agitation. The disrupted cells were diluted by adding 0.5 ml TE to each tube; this was done to allow phase separation after treatment with phenol in the following step. Deproteinization was carried out by treatment with 1 volume of liquid phenol equilibrated with TE. The nucleic acids in the aqueous phase were treated with ether to eliminate residual phenol, and recovered by isopropanol was eliminated by treatment (DNase ,tg/4tl, ,tg