Legume Research

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Legume Research, volume 40 issue 4 (august 2017) : 643-648

Determination of Cry1Ac copy number in transgenic pigeonpea plants using quantitative real time PCR

Meenakshi Jain, Surender Khatodia, Pushpa Kharb, Parveen Batra, Vijay K. Chowdhury
1<p>Department of Molecular Biology and Biotechnology,&nbsp;CCS Haryana Agricultural University, Hisar- 125 004, India.</p>
Cite article:- Jain Meenakshi, Khatodia Surender, Kharb Pushpa, Batra Parveen, Chowdhury K. Vijay (2016). Determination of Cry1Ac copy number in transgenic pigeonpeaplants using quantitative real time PCR . Legume Research. 40(4): 643-648. doi: 10.18805/lr.v0iOF.11300.

Copy number of Cry1Ac in transgenic pigeonpea plants was determined by quantitative real time PCR using Syber Green as fluorescence indicator. Gene specific primers designed to amplify relatively long amplicons (400 – 600 bp), for Cry1Ac was used to increase specificity and sensitivity of Real time PCR. Estimated copy number in transgenic lines using real-time quantitative PCR and southern hybridization was correlated and found to be same i.e. single copy number. This study shows effectivness of real-time PCR method for estimating the transgene copy number in transgenic pigeonpea plants by a simple, accurate and cost effective manner.

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