title: An extremely simple and effective colony PCR procedure for bacteria, yeasts, and microalgae.
abstract: An extremely simple and effective colony PCR procedure is established for both gram-negative and gram-positive bacteria, yeasts, and microalgae. Among the four lysis buffers examined, Y-PER is observed to be more effective than Tris/EDTA, 0.2 % SDS, and 10 mM EDTA in the extraction of PCR-quality genomic DNA from those microorganisms. Vortexing or pipetting agitation of the cells in Y-PER for 5-10 s was sufficient to release genomic DNA for all the test bacteria and yeasts, and most microalgae. Additional incubation at 98 °C for 5 min for further cell disruption was essential only for Chlorella vulgaris due to its notoriously rigid cell wall.
authors.affiliation: Mathematisch-Naturwissenschaftliche Fakultät, Rheinische Friedrich-Wilhelms Universität Bonn, Wegelerstraße 10, 53115 Bonn, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: Packeiser
authors.forename: Chanyuen
authors.initials: C
authors.lastname: Lim
authors.forename: Balaji
authors.initials: B
authors.lastname: Balagurunathan
authors.forename: Jinchuan
authors.initials: J
authors.lastname: Wu
authors.forename: Hua
authors.initials: H
authors.lastname: Zhao
mesh_terms: D002458:Cell Fractionation / Q000379:methods*; D048028:DNA, Algal / Q000235:genetics* / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D004271:DNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D058086:Microalgae / Q000235:genetics*; D059013:Microbial Consortia / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D048028:DNA, Algal; D004269:DNA, Bacterial; D004271:DNA, Fungal