Referral Center for Molecular Diagnosis of Transgenic Planting Materials

Delhi, India

Referral Center for Molecular Diagnosis of Transgenic Planting Materials

Delhi, India
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Randhawa G.J.,Referral Center for Molecular Diagnosis of Transgenic Planting Materials | Singh M.,Referral Center for Molecular Diagnosis of Transgenic Planting Materials | Chhabra R.,Referral Center for Molecular Diagnosis of Transgenic Planting Materials | Sharma R.,Referral Center for Molecular Diagnosis of Transgenic Planting Materials
Food Analytical Methods | Year: 2010

Qualitative and quantitative PCR assays were developed for detection of commercialised Bt cotton events, i. e. MON531, MON15985 and other Bt crops, which are under different stages of field trials in India, i. e. Bt brinjal, Bt rice, Bt cauliflower, Bt potato and Bt okra. Multiplex PCR assays simultaneously detecting specific cry1Ac, cry1Ab, cry2Ab genes, Cauliflower Mosaic Virus 35S promoter, nptII marker gene along with species- or taxon-specific endogenous gene in these Bt crops have been developed. The quantitative real-time PCR assays were also reported for cry1Ac gene using designed primers and TaqMan probe. The sensitivity of developed assays for detection of specific transgene was established up to 0.01%. The analytical methods developed in the present report will be of immense use for qualitative screening and detection of Bt crops along with the quantitative analysis of inserted cry1Ac gene to meet the threshold level for regulatory compliance. © 2010 Springer Science+Business Media, LLC.


PubMed | Referral Center for Molecular Diagnosis of Transgenic Planting Materials
Type: Comparative Study | Journal: Journal of agricultural and food chemistry | Year: 2010

The genetically modified (GM) Bt crops expressing delta-endotoxins from Bacillus thuringiensis provide protection against a wide range of lepidopteron insect pests throughout the growing season of the plant. Bt cotton is the only commercialized crop in India that is planted on an area of 7.6 million hectares. With the increase in development and commercialization of transgenic crops, it is necessary to develop appropriate qualitative and quantitative methods for detection of different transgenic events. The present study reports on the development of a decaplex polymerase chain reaction (PCR) assay for simultaneous detection of transgene sequences, specific transgene constructs, and endogenous stearoyl acyl desaturase (Sad1) gene in two events of Bt cotton, i.e., MON531 and MON15985. The decaplex PCR assay is an efficient tool to identify and discriminate the two major commercialized events of Bt cotton, i.e., MON531 and MON15985, in India. Real-time PCR assays were also developed for quantification of cry1Ac and cry2Ab genes being employed in these two events. The quantitative method was developed using seven serial dilutions containing different levels of Bt cotton DNA mixed with a non-Bt counterpart ranging from 0.01 to 100%. The results revealed that the biases from the true value and the relative standard deviations were all within the range of 20%. The limit of quantification (LOQ) of the developed real-time PCR method has also been established up to 0.01%.


Randhawa G.J.,Referral Center for Molecular Diagnosis of Transgenic Planting Materials | Chhabra R.,Referral Center for Molecular Diagnosis of Transgenic Planting Materials | Singh M.,Referral Center for Molecular Diagnosis of Transgenic Planting Materials
Journal of Agricultural and Food Chemistry | Year: 2010

The genetically modified (GM) Bt crops expressing delta-endotoxins from Bacillus thuringiensis provide protection against a wide range of lepidopteron insect pests throughout the growing season of the plant. Bt cotton is the only commercialized crop in India that is planted on an area of 7.6 million hectares. With the increase in development and commercialization of transgenic crops, it is necessary to develop appropriate qualitative and quantitative methods for detection of different transgenic events. The present study reports on the development of a decaplex polymerase chain reaction (PCR) assay for simultaneous detection of transgene sequences, specific transgene constructs, and endogenous stearoyl acyl desaturase (Sad1) gene in two events of Bt cotton, i.e., MON531 and MON15985. The decaplex PCR assay is an efficient tool to identify and discriminate the two major commercialized events of Bt cotton, i.e., MON531 and MON15985, in India. Real-time PCR assays were also developed for quantification of cry1Ac and cry2Ab genes being employed in these two events. The quantitative method was developed using seven serial dilutions containing different levels of Bt cotton DNA mixed with a non-Bt counterpart ranging from 0.01 to 100%. The results revealed that the biases from the true value and the relative standard deviations were all within the range of ±20%. The limit of quantification (LOQ) of the developed real-time PCR method has also been established up to 0.01%. © 2010 American Chemical Society.

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