Rapid determination of genomic DNA length for new bacteriophages.
dsDNA viruses with long genomes (>200 kb) are expected to be a major source of novel genes. To rapidly characterize the genomes of newly isolated dsDNA bacteriophages, we develop here a procedure for the PFGE of intact long DNA genomes from bacteriophage particles in unfractionated, infected cell lysates of either liquid or gelled cultures. The DNA used for PFGE is suitable for sequencing after extraction with phenol. The PFGE is tuned to the range of expected DNA lengths. This procedure bypasses the isolation of bacteriophage particles and is useful for PFGE analysis of DNA from dissected zones of bacteriophage plaques.
Serwer
Philip
P
Department of Biochemistry, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229-3900, USA. serwer@uthscsa.edu
Hayes
Shirley J
SJ
Thomas
Julie A
JA
Griess
Gary A
GA
Hardies
Stephen C
SC
D001435:Bacteriophages / Q000235:genetics* / Q000648:ultrastructure; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods; D004783:Environmental Microbiology; D019143:Evolution, Molecular; D016679:Genome, Viral; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D011387:Prokaryotic Cells / Q000821:virology*
D004279:DNA, Viral; D004262:DNA Restriction Enzymes