An optimized affordable DNA-extraction method from Salmonella enterica Enteritidis for PCR experiments

Ghaderi, Rainak; Moradi Bidhendi, S.; Khaki, P.; Tadayon, K.; Karimnasab, Nasim; Sekhavati, M.; Asadi, F.

doi:10.7508/ari.2013.02.003

An optimized affordable DNA-extraction method from Salmonella enterica Enteritidis for PCR experiments

Authors

¹ Razi vaccine & Serum Research Institite AND Armenian National Agrarian University, Yerevan, Armenia

² Razi Vaccine & Serum Research Institute

³ Razi vaccine & Serum Research Institite AND Azad University-Karaj Branch

⁴ Department of Animal Science, Ferdowsi University of Mashhad, Mashhad, Iran

10.7508/ari.2013.02.003

Abstract

In diagnostic and research bacteriology settings with budget and staff restrictions, fast and cost-effective genome extraction methods are desirable. If not inactivated properly, cellular and/or environmental DNA nucleases will degrade genomic material during the extraction stage, and therefore might give rise to incorrect results in PCR experiments. When crude cell extracts, proteinase K–treated templates and purified DNAs prepared by phenol-chloroform-isoamylalcohol method as well as a commercial extraction kit were subjected to the Salmonella enterica Enteritidis specific STM2 PCR, with exception of crude cell extract, PCR products from all other three methods saved their integrity for 28 days post-generation. This work aimed to find out whether improvement to boiling method can guaranty stability of PCR products. As results showed, treatment of crude cell extracts from S. Enteritidis with proteinase K offers an inexpensive, fast and effective DNA extraction method suitable for high-throughput laboratories.

Keywords