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A rapid and cost–efficient DMSO–based method for isolating DNA from cultured lichen photobionts
Author(s) -
del Campo Eva M.,
Hoyo Alicia del,
Casano Leonardo M.,
Martínez-Alberola Fernando,
Barreno Eva
Publication year - 2010
Publication title -
taxon
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 81
eISSN - 1996-8175
pISSN - 0040-0262
DOI - 10.1002/tax.592023
Subject(s) - dimethyl sulfoxide , dna extraction , dna , centrifugation , extraction (chemistry) , chromatography , phenol , solvent , contamination , chemistry , biochemistry , biology , polymerase chain reaction , organic chemistry , ecology , gene
We have developed a simple and fast procedure for the purification of PCR–quality DNA from cultured lichen photobionts. This new one–step method uses the solvent dimethyl sulfoxide (DMSO) combined with heat treatment to chemically breakdown algal and plant tissues. The DMSO–DNA extracts may be directly precipitated and purified by standard techniques in a total time of approximately 30 min. Compared to other DNA extraction protocols, the DMSO–based method suppresses the need for liquid nitrogen, any extraction buffer, grinding, phenol, and long incubation or centrifugation steps, thereby considerably reducing the possibility of contamination. In addition, minimal amounts of starting material (5 × 106–20 × 106 cells from liquid or agarized cultures) produce sufficient DNA for 200 PCR reactions approximately, making this protocol a practical option for colony screening. This method works well in a wide range of cultured lichen photobionts and reduces the amount of labor–intensive steps and time consumed by other multi–step procedures, allowing for efficient processing of an increased number of samples.