Use of polymerase chain reaction (PCR) fingerprinting to differentiate bacteria for microbial products screening

Abstract

PCR fingerprinting offers a practical molecular means to quickly and reliably differentiate bacteria for microbial products screening. A combination of low resolution and high resolution PCR fingerprinting provides a hirarchical system which allows the discrimination of bacteria at species and subspecies level within 7 h. DNA was extracted from cells by incubating them in water at 95°C for 30 min. A sample of 1 μl of the cell-free aqueous extract then was used as a source of template DNA in the PCR. The PCR products were separated by electrophoresis on an acrylamide gel and visualized by ethidium bromide staining. The band patterns generated for each different culture were unique, reproducible, and independent of cultivation conditions. Band patterns may be compared visually or by using imaging and pattern matching software. In our laboratory, bacteria such as actinomycetes, Gram-negative and Gram-positive soil eubacteria, and photosynthetic non-sulfur bacteria have been differentiated using PCR fingerprinting.