异戊醇
溶解
裂解缓冲液
十二烷基硫酸钠
DNA
基因组DNA
色谱法
生物
碱裂解
苯酚萃取
化学
DNA提取
生物化学
蛋白酶K
核糖核酸
分离(微生物学)
分子生物学
细菌
核酸
滚动圆复制
基因
聚合酶链反应
酒
质粒
遗传学
dna疫苗
作者
Debabrata Majumdar,Yael J. Avissar,James H. Wyche
出处
期刊:PubMed
日期:1991-07-01
卷期号:11 (1): 94-101
被引量:35
摘要
A very simple procedure for the simultaneous preparation of genomic DNA and total RNA is described. The procedure is essentially the same for eukaryotes and prokaryotes except for the lysis buffer and can be used for small or large numbers of cells. Mammalian cells are lysed in sodium dodecyl sulfate and bacterial cells are lysed in Triton X-100, both in the presence of EDTA. RNA is obtained in the aqueous phase after phenol (acidic pH):chloroform:isoamyl alcohol extraction. DNA is eluted out of the organic phase (and the interface) into the aqueous phase by increasing the pH with highly basic 1 M Tris solution. The method is extremely rapid for small or large numbers of cells, and several large samples can be processed in one day. The qualities of both nucleic acids are excellent and the yield is high.
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