番茄叶片基因组DNA快速制备技术及其在基于实时荧光定量PCR的转基因检测中的应用

doi:10.3724/SP.J.1005.2011.01017

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HEREDITAS ›› 2011, Vol. 33 ›› Issue (9): 1017-1022.doi: 10.3724/SP.J.1005.2011.01017

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Techniques for rapid preparation of tomato leaf DNA and its application in real-time quantitative PCR-based transgene detection

WANG Wei-Wei, ZHU Chang-Qing, LIU Xiao-Hua, CHEN Kun-Song, XU Chang-Jie

Department of Horticulture, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China

Received:2010-12-21 Revised:2011-05-18 Online:2011-09-20 Published:2011-09-25

Abstract

Abstract: Using tomato (Solanum lycopersicum L. cv. Micro-Tom) leaf as material, a simple and rapid DNA preparation protocol was established. This method required only 2-20 mm2 leaf with only one extraction solution and involved one pipetation and one centrifugation each. No precipitation was required. The suitable volume of prepared DNA solution, as PCR template, for real-time quantitative PCR was determined to be 0.1-0.2 mL in 12.5 mL final reaction volume. The ex-cessive template DNA solution was confirmed to reduce PCR efficiency and even can result in PCR failure. This technique for rapid preparation of DNA and a compatible real-time quantitative PCR were successfully applied in transgene detection of tomato plants.

Key words: tomato, rapid DNA preparation, real-time quantitative PCR, transgene detection

WANG Wei-Wei, SHU Chang-Jing, LIU Xiao-Hua, CHEN Hun-Song, XU Chang-Jie. Techniques for rapid preparation of tomato leaf DNA and its application in real-time quantitative PCR-based transgene detection[J]. HEREDITAS, 2011, 33(9): 1017-1022.

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Techniques for rapid preparation of tomato leaf DNA and its application in real-time quantitative PCR-based transgene detection

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