Selection of Universal Primers for PCR Quantification of Total Bacteria Associated With Fish Fillets

Abstract

An optimal universal primer pair DG74/RW01 for quantitative PCR detection of the mixed bacterial flora from fish fillets was selected from five pairs of primers. The minimum detection limits with GelStar™ and ethidium bromide (EB) used as agarose stains for DNA bands with these primers were found to be 5 and 1 × 10² CFU/PCR, respectively. The liner range of DNA amplification was from 50 to 1 × 10⁵ with GelStar™ and from 5 × 10² to 1 × 10⁵ with EB.

Key Words:

• universal primers
• fish fillets
• PCR

ACKNOWLEDGEMENTS

Jung-Lim Lee was supported in part by a postdoctoral fellowship grant from the Korea Research Foundation funded by the government of Korea (MOEHRD, Basic Research Promotion Fund, M01-2004-000-20192-0) and by a U.S.D.A Special Seafood Safety grant. This is paper no. 3363 from the Massachusetts Agricultural Experiment Station, University of Massachusetts, Amherst, Mass., USA.