A new approach to the isolation of genomic DNA from yeast and fungi: Preparation of DNA-containing cell envelopes and their use in PCR

被引:7
作者
Danilevich, VN [1 ]
Grishin, EV [1 ]
机构
[1] Russian Acad Sci, Shemyakin Ovchinnikov Inst Bioorgan Chem, Moscow 117997, Russia
关键词
DNA-containing cell envelopes; genomic DNA isolation; PCR amplification; protein and lipid extraction; yeast and filalmentous fungi;
D O I
10.1023/A:1015021508321
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A simple and rapid procedure for the preparation of yeast and fungal DNA samples useful in PCR amplification was developed. The DNA was purified from proteins, lipids, polysaccharides, and other impurities by high-temperature extraction (in a boiling water bath) with buffer solutions containing chaotropic salts. Under these conditions, yeast and fungal cell envelopes remain unbroken and retain the original DNA and RNA that could be used for direct PCR amplification. We called the proposed PCR technique as the PCR using DNA-containing cell envelopes.
引用
收藏
页码:136 / 146
页数:11
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