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A calibrator plasmid for quantitative analysis of insect resistant maize (Yieldgard MON 810.

Rajashekhar, V. Ballari and Asha, Martin and Lalitha, R. Gowda (2013) A calibrator plasmid for quantitative analysis of insect resistant maize (Yieldgard MON 810. Food Chemistry, 140. pp. 382-389. ISSN 0308-8146

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Abstract

Real-time PCR (RT-PCR) is the preferred method for the quantification of genetically modified organisms (GMOs) and implementation of labeling regulations. The precision, sensitivity, and reproducibility of RTPCR data depend on the use of external calibrators. In this investigation, a dual target plasmid designated pRSETMON-02 comprising of MON 810 maize event specific and endogenous zein gene sequences in 1:1 ratio in tandem was constructed and validated. Commutability of plasmid DNA (pDNA) and genomic DNA (gDNA) calibrators for the quantification of MON 810 maize was assessed by employing a TaqMan RT-PCR targeting the P-35S and zein gene. Higher PCR efficiencies, good linearity and lower relative standard deviation (RSD) values were associated with pRSETMON-02 as opposed to gDNA calibrants. pDNA calibrants exhibited better performance characteristic in terms of closeness to the expected value of unknown samples than their genomic counterparts. Short term stability study of the pRSETMON-02 plasmid stored at different temperatures showed that pDNA is stable for 45 days at �20, and 4 �C. The results demonstrated that the developed dual target plasmid pRSETMON-02 is fit for the intended use of quantifying MON 810 maize and is a better alternative to conventional seed powder calibrants.

Item Type: Article
Uncontrolled Keywords: Genetically modified organisms MON 810 maize Real-time PCR Reference materials Plasmid DNA calibrants Genomic DNA calibrants
Subjects: 600 Technology > 08 Food technology > 22 Legumes-Pulses > 02 Maize
Divisions: Food Safety Analytical Quality Control Lab
Protein Chemistry and Technology
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 26 Mar 2013 06:02
Last Modified: 28 Jun 2016 08:28
URI: http://ir.cftri.com/id/eprint/11163

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