Detecting cry1Ac by loop mediated isothermal amplification by SYBR green-I

Abstract

Detection of Genetically Modified Organisms (GMOs) is an important part of GMO labelling, as without detection methods the traceability of GMOs would rely solely on documentation. Efficient detection strategies for GM crops need to be in compliance with regulatory frameworks and address consumer concerns. The DNA based techniques are currently the major detection methods that are widely used due to their ease and accuracy Loop mediated isothermal amplification (LAMP) is a simple, rapid, specific and cost effective nucleic acid amplification method. This method employs a DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA. A set of six specific primers was designed to recognize six distinct sequences on the target cry1Ac gene, including a pair of inner primers, a pair of outer primers, and a pair of loop primers. The optimum reaction temperature and time were optimised to be 65°C for 60 min respectively and stained with SYBR Green –I for visual detection of GMO.