Legume Research-An International Journal

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

DOI: 10.18805/lr.v0iOF.11300    | Article Id: LR-3675 | Page : 643-648
Citation :- Determination of Cry1Ac copy number in transgenic pigeonpeaplants using quantitative real time PCR .Legume Research-An International Journal.2017.(40):643-648

Meenakshi Jain, Surender Khatodia, Pushpa Kharb, Parveen Batra and Vijay K. Chowdhury

Meenakshi110@gmail.com
Address :

Department of Molecular Biology and Biotechnology, CCS Haryana Agricultural University, Hisar- 125 004, India.

Submitted Date : 30-12-2015
Accepted Date : 23-04-2016
First Online:

Abstract

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.

Keywords

Copy number Cry1Ac Pigeonpea transformation Real-time PCR Syber green.

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