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To this effect, 340 clinical, veterinary, and environmental isolates from Argentina, Brazil, Chile, Colombia, Mexico, Peru, Venezuela, Guatemala, and Spain were typed by using M13 polymerase chain reaction-fingerprinting and orotidine monophosphate pyrophosphorylase (URA5) gene restriction fragment length polymorphsm analysis with Hhal and Sau961 in a double digest.
All 340 isolates were typed by PCR fingerprinting by using the minisatellite-specific oligonucleotide MI3 as a single primer and RFLP analysis of the URA5 gene with the restriction enzymes Sau96I and HhaI in a double digest.
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Restriction sites of 10 restriction endonucleases were mapped in the two PCR-amplified mtDNA regions by using double digests.
To answer these questions, we conducted restriction site analyses on five individuals from each of 15 different Sebastes species common in Alaskan waters and mapped the sites using double digests to determine individual-based haplotypes.
Double digests were examined both in agarose and polyacrylamide by using 100- and 25-bp ladders.