Anwaar Ahmad
Department of Crop and Soil Sciences, 361 Plant and Soil Science Building, USA
Shahina Bano Maqbool
Department of Crop and Soil Sciences, 361 Plant and Soil Science Building, USA
Syed Anwar Hashsham
Department of Civil and Environmental Engineering, Al6 Engineering Research Complex,
Michigan State University, East Lansing, MI 48824, USA
Mariam B. Sticklen
Department of Crop and Soil Sciences, 361 Plant and Soil Science Building, USA
ABSTRACT
We have developed a simple real-time PCR procedure for determination of transgene integration and copy numbers. Transgenic rice plants harboring cry1Ab and cry1Ac were produced via Agrobacterium- mediated transformation. Gene specific primers designed to amplify a relatively long amplicon, 542 bp for cry1Ab and 606 bp for cry1Ac were used to increase the specificity and sensitivity of the real-time PCR. The assay was carried out using the ABI Prism 7700 sequence detection system with Syber Green I as the fluorescence indicator. Using real-time PCR, we calculated 1-3 copies of transgenes (cry1Ab/cry1Ac) in our transgenic rice lines. The accuracy of real-time PCR results was determined with results obtained after Southern blot analyses. Comparison showed 75-100% accuracy of our Syber Green I real-time PCR with Southern blots.
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How to cite this article
Anwaar Ahmad, Shahina Bano Maqbool, Syed Anwar Hashsham and Mariam B. Sticklen, 2005. Determination of cryIAb and cryIAc Copy Number in Transgenic
Basmati 370 Rice (Oryza sativa L.) Plants Using Real-time PCR and its
Comparison with Southern Blot. Journal of Biological Sciences, 5: 283-288.
DOI: 10.3923/jbs.2005.283.288
URL: https://scialert.net/abstract/?doi=jbs.2005.283.288
DOI: 10.3923/jbs.2005.283.288
URL: https://scialert.net/abstract/?doi=jbs.2005.283.288
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