BMB Reports
- Volume 33 Issue 6
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- Pages.537-540
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- 2000
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- 1976-670X(eISSN)
A Simple and Reliable Method for Preparation of Cross-Contamination-Free Plant Genomic DNA for PCR-Based Detection of Transgenes
- Hwang, Seon-Kap (GMO Safety Assessment Team, Division of Biochemistry, National Institute of Agricultural Science and Technology (NIAST), Rural Development Administration (RDA)) ;
- Kim, Young-Mi (GMO Safety Assessment Team, Division of Biochemistry, National Institute of Agricultural Science and Technology (NIAST), Rural Development Administration (RDA))
- Received : 2000.08.17
- Accepted : 2000.10.01
- Published : 2000.11.30
Abstract
A simplified but reliable method was developed for the polymerase chain reaction (PCR)-based detection of genetically modified (GM) plants. The modified CTAB (mCTAB) method enabled us to prepare a high quality of genomic DNA from several hundred plant leaf samples in one day. Using DNA samples prepared from seven dicots and two monocots, approximately 1.75-kb regions spanning 17 S to 25 S ribosomal RNA genes were successfully amplified in a 2X PCR pre-mix containing BLOTTO. Further fidelity assessment of the mCTAB method by PCR analysis with Roundup Ready soybean (RRS) and non-RRS plants showed that the DNA samples prepared alternately from each of two lines were evidently free of cross-contamination. These results demonstrate that the mCTAB method is highly recommended for the rapid detection of transgenes in large numbers of leaf samples from diverse transgenic plants.