FOOD SCIENCE ›› 2014, Vol. 35 ›› Issue (2): 243-247.doi: 10.7506/spkx1002-6630-201402047

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Pretreatment Optimization of Simulated Rice Cake Samples for Fluorescence Polymerase Chain Reaction to Detect Transgenic Components

ZHANG Bing1,2, WANG Zhi-chun3, SHAO Bi-ying1, MIAO Ting-yu1, PENG Juan1, CHEN Wen-bing1,*   

  1. 1. Inspection and Quarantine Techonology Center, Fujian Entry-Exit Inspection and Quarantine Bureau, Fuzhou 350003, China;
    2. Xiamen Tasman Bio-Tech Co. Ltd., Xiamen 361023, China; 3. Fujian Provincial Seed Station, Fuzhou 350003, China)
  • Received:2013-07-23 Revised:2013-12-18 Online:2014-01-25 Published:2014-02-19
  • Contact: CHEN Wen-bing E-mail:1273495290@qq.com

Abstract:

In order to improve the sensitivity and detection limits of transgenic components in rice products, pretreatment
conditions for DNA extraction including sample particle size and hexadecyltrimethylammonium bromide (CTAB) lysis
buffer were optimized using simulated transgenic rice cake samples. The results showed that the content of DNA had
significant differences among 9 treatment combinations of sample particle size and CTAB lysis buffer incubation time. The
content of DNA extracted by the combination of sample particle size larger than 100 mesh and CTAB lysis buffer incubation
time longer than 8 h (overnight) was the highest. The detection limit of major transgenic components in rice cake samples
was reduced from transgenic content of 0.01% (m/m) to 0.001% (m/m), suggesting that the sensitivity of fluorescence
polymerase chain reaction (PCR) was improved by 10 times using the optimal pretreatment. The detection accuracy of
genetically modified components in rice products was effectively improved as well.

Key words: rice products (rice cake), genetically modified ingredients, fluorescence real time-polymerase chain reaction (RT-PCR), limit of detection

CLC Number: