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Appl Biochem Biotechnol ; 169(2): 695-700, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23271627

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.


Subject(s)
Cell Fractionation/methods , DNA, Algal/genetics , DNA, Bacterial/genetics , DNA, Fungal/genetics , Microalgae/genetics , Microbial Consortia/genetics , Polymerase Chain Reaction/methods , DNA, Algal/isolation & purification , DNA, Bacterial/isolation & purification , DNA, Fungal/isolation & purification
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