A general method of isolating high molecular weight DNA from methanogenic archaea (archaebacteria).
High molecular weight DNA was readily isolated from all methanogens treated, as well as from thermophilic anaerobic eubacteria, by grinding cells frozen in liquid N2, prior to lysis with SDS. DNA can subsequently be purified by the usual phenol-chloroform extractions. The procedure yields DNA readily cut by restriction enzymes and suitable for oligonucleotide probing, as well as for mole percent G + C content determination by thermal denaturation. The method routinely yields DNA of high molecular weight and is an improvement over DNA isolation methods for many methanogens, which often involve an initial breakage of the cells in a French pressure cell.
Jarrell
K F
KF
Department of Microbiology and Immunology, Queen's University, Kingston, Ont., Canada.
Faguy
D
D
Hebert
A M
AM
Kalmokoff
M L
ML
D001105:Archaea / Q000737:chemistry; D001431:Bacteriological Techniques; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification*; D019605:Euryarchaeota / Q000737:chemistry*; D008970:Molecular Weight
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes