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Detection and quantification of Streptomyces violaceolatus plasmid DNA in soil
Author(s) -
Cresswell N.,
Saunders V.A.,
Wellington E.M.H.
Publication year - 1991
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.1991.tb00606.x
Subject(s) - lysis , spore , plasmid , dna extraction , dna , alkaline lysis , biology , mycelium , microbiology and biotechnology , streptomyces , extraction (chemistry) , southern blot , inoculation , plasmid preparation , bacteria , chromatography , chemistry , botany , polymerase chain reaction , horticulture , biochemistry , genetics , gene , dna vaccination , pbr322
Two methods for the direct extraction of DNA from soil were investigated using soil inoculated with Streptomyces violaceolatus ISP5438 harbouring the multicopy plasmid pIJ673. Detection limits for plasmid DNA were determined by Southern blot technique. An SDS/heat lysis method gave approximately two orders of magnitude less sensitivity than lysis and extraction by bead‐beating soil inoculated with spores. The use of these two methods allowed differentiation between spore‐ and mycelial‐borne DNA. This was due to the resistance of the spores to lysis when subjected to SDS/heat lysis.