Russian Journal of Bioorganic Chemistry, Vol. 28, No. 2, 2002, đ . 136
A New Approach to the Isolation of Genomic DNA from Yeast and Fungi: Preparation of DNA-containing Cell Envelopes and Their Use in PCR
V. N. Danilevich* , 1 and E. V. Grishin
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, GSP-7 Moscow , 117997 Russia
Abstract: 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.
Key words: DNA-containing cell envelopes, genomic DNA isolation, PCR amplification, protein and lipid extraction, yeast and filamentous fungi