Improved PCR-based method for the direct screening of Streptomyces transformants.
Streptomycetes are attractive microorganisms because of their high secretion capacity and for the production of secondary metabolites. We report the improvement of a PCR-based method for screening of solid media-grown Streptomyces transformants. By resuspending mycelium into 2.5-10% dimethyl sulfoxide (DMSO), extensive manipulation prior to PCR could be avoided, thus significantly reducing screening time. Results proved to be both reliable and specific.
Van Dessel
W
W
Laboratory of Bacteriology, Rega Institute, Katholieke Universiteit Leuven, Minderbroedersstraat 10, B-3000 Louvain, Belgium.
Van Mellaert
L
L
Geukens
N
N
Anné
J
J
D004121:Dimethyl Sulfoxide / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013302:Streptomyces / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D014169:Transformation, Bacterial
D004121:Dimethyl Sulfoxide