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Extraction of DNA from Phytophthora infestans Using QIAGEN Columns
Author(s) -
Tooley P. W.,
Carras M. M.
Publication year - 1996
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/j.1439-0434.1996.tb00308.x
Subject(s) - phytophthora infestans , biology , mycelium , dna extraction , dna , chromatography , extraction (chemistry) , genomic dna , phytophthora , microbiology and biotechnology , botany , blight , genetics , polymerase chain reaction , gene , chemistry
High molecular weight DNA of Phytophthora infestans was extracted using a modification of the commercial QIAGEN column procedure. Both a ‘maxi’ and ‘mini’ procedure are described. The maxi procedure utilizes a QIAGEN‐tip 500 column while the mini procedure utilizes a QIAGEN‐tip 20 column. When fungal protoplasts were used as starting material from 9 g (fresh weight) of mycelium, nearly 500 μg of DNA in the size range of 20–200 kb was obtained and the product was successfully used in construction of a lambda genomic library. The modified QIAGBN method can replace the more timeconsuming, and expensive cesium chloride density gradient centrifugation for extraction of ultra‐pure DNA from P. infestans .