Optimization of RAPD PCR Assay for Salmonella Detection

doi:10.7506/spkx1002-6300-200920070

FOOD SCIENCE ›› 2009, Vol. 30 ›› Issue (20): 320-323.doi: 10.7506/spkx1002-6300-200920070

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Optimization of RAPD PCR Assay for Salmonella Detection

SU Wan-yuan1，HOU Jia-kui2，LIU Ting1，GUO Ai-ling1,*，WANG Hong-jiang1，XIE Ji1，MA Mei-hu1

：TS201.31. College of Food Science and Technology, Huazhong Agricultural University,Wuhan 430070, China ；
2. Xuchang Vocational Technical College, Xuchang 461000, China

Received:2008-10-27 Revised:2009-07-28 Online:2009-10-15 Published:2010-12-29
Contact: GUO Ai-ling E-mail:guoailing@mail.hzau.edu.cn

Abstract

Abstract:

A random amplification of polymorphic DNA (RAPD)-PCR assay was developed for detecting Salmonella using a primer of 5′-CCGAAGCTGC-3′. In order to obtained distinctive and reliable results, the optimal PCR reaction conditions (namely PCR system volume, renaturation temperature, primer concentration and Mg2+ concentration) and agarose concentration for electrophoresis were investigated. Results showed that the optimal RAPD-PCR reaction was performed in a 25μl system containing 2.0μl of 25 mmol/L MgCl2 and 0.2 μl of primer at 32 ℃ and the optimal agarose concentration was 1.0%. Under such conditions, sufficient and clear electrophoresis strips were observed in RAPD-PCR detection of 3 typical Salmonella strains and 9 isolated Salmonella strains.

Key words: Salmonella, local strains, RAPD, optimization

CLC Number:

TS201.3

SU Wan-yuan1，HOU Jia-kui2，LIU Ting1，GUO Ai-ling1,*，WANG Hong-jiang1，XIE Ji1，MA Mei-hu1. Optimization of RAPD PCR Assay for Salmonella Detection[J]. FOOD SCIENCE, 2009, 30(20): 320-323.

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Optimization of RAPD PCR Assay for Salmonella Detection

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