Rapid PCR Fingerprinting of Bacterial Genomes with REP Primers in Capillary Tubes Using the Air Thermo-Cycler.

R. A. DEWEY; O GRAU; A LAGARES

The RapidCyclist

Idaho Technologies

Año: 1996 vol. 3 p. 10 - 11

DNA fingerprinting of genomes using PCR methods has been intensively used during the last years to characterize genomic diversity and to search for specific DNA markers (1-9). Different DNA banding patterns have been obtained using primers with length ranging from 8 to 25 nucleotides containing either arbitrary or specific sequences. In particular, the use of bacterial Repetitive Extragenic Palindromic sequences (REP) and Enterobacterial Repetitive Intergeneric Consensus (ERIC) have been proved to be practical and appropriate to fingerprint a number of different bacterial species. Although REP and ERIC primers do not lead to amplification patterns as complex as those obtained with the random primed DAF, visualization of amplified DNA fragments can be easily achieved by agarose gel electrophoresis/ethidium bromide staining. Thus, classic REP and ERIC PCR amplifications may be efficiently used to characterize Gram-negative and Gram-positive bacterial genomes in a 10 hours experimental procedure. Here, we report a simple, rapid and  reproducible protocol to perform REP DNA amplifications in 2 h using capillary tubes in air thermo-cyclers.