~ 2176 ~ Journal of Pharmacognosy and Phytochemistry 2018; 7(2): 2176-2180 E-ISSN: 2278-4136 P-ISSN: 2349-8234 JPP 2018; 7(2): 2176-2180 Received: 17-01-2018 Accepted: 18-02-2018 Preeti Jambagi Department of Molecular Biology and Agricultural Biotechnology University of Agricultural Sciences, Raichur, Karnataka, India Kisan B Department of Molecular Biology and Agricultural Biotechnology University of Agricultural Sciences, Raichur, Karnataka, India Muralidhar B Department of Molecular Biology and Agricultural Biotechnology University of Agricultural Sciences, Raichur, Karnataka, India Shruthi Nagaral K Department of Molecular Biology and Agricultural Biotechnology University of Agricultural Sciences, Raichur, Karnataka, India I Shankergoud Department of Molecular Biology and Agricultural Biotechnology University of Agricultural Sciences, Raichur, Karnataka, India JM Nidagundi Department of Molecular Biology and Agricultural Biotechnology University of Agricultural Sciences, Raichur, Karnataka, India Ayyangouda Patil Department of Molecular Biology and Agricultural Biotechnology University of Agricultural Sciences, Raichur, Karnataka, India Correspondence Preeti Jambagi Department of Molecular Biology and Agricultural Biotechnology University of Agricultural Sciences, Raichur, Karnataka, India Detecting cry1Ac by loop mediated isothermal amplification by SYBR green-I Preeti Jambagi, Kisan B, Muralidhar B, Shruthi Nagaral K, I Shankergoud, JM Nidagundi and Ayyangouda Patil 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. Keywords: LAMP, GMO, detection Introduction Management of insect pests through synthetic insecticides is being practiced and was a boon in the advent era of green revolution. Sole reliance on insecticides has caused an imbalance in the agro-ecosystem creating resistance and resurgence problems warranting alternate control measures. As an alternative to the insecticides inserting of foreign gene through genetic engineering and evolving transgenic cotton is considered to be an important strategy for the management of major pests of cotton, particularly bollworms without inimical effects on ecosystem. Bt cotton was among the first GM crops to be commercialised during the 1900s at the global level and was officially approved for sale in India in 2002. It consists of a gene from the soil bacterium Bacillus thuringensis (Bt), which provides resistance to different bollworm species, a major pest in cotton and helps cotton growers benefit through efficient pest control. India is the second largest producer and consumer of cotton accounting 33℅of the global cotton production in 2015 - 16 (Anon) and is cultivated in an area of about 11.8mha. However consumer concerns related to development and marketing of GMO and derived food products have resulted in increased awareness regarding the food labelling all over the world. The development and commercialisation of GM crops is increasing at a faster pace, to develop qualitative and quantitative methods for detection of GM crops has become even more challenging. Polymerase Chain Reaction (PCR) being a reliable, robust and sensitive technique, has broad application in GM detection. However, high-precision equipment and procedures associated with PCR analysis are some of the constraints, which limit their use for on-site detection. Moreover, the qPCR technique is often sensitive to inhibitors present in plant extracts. [Boonham et al] [2] Loop-mediated isothermal amplification (LAMP), an isothermal nucleic acid amplification technique, is less sensitive to inhibitors, does not require sophisticated equipment, and has the potential to be deployed on site. [Notomi et al., 2002] [3] LAMP is gaining importance because of its sensitivity and specificity, being superior to PCR and usually comparable to qPCR. In addition, LAMP provides results on site in a significantly shorter time than conventional PCR- based techniques. LAMP is characterized by the use of four different primers, specifically designed to recognize six distinct regions on the target DNA template. An inner primer containing sequences of sense and antisense strands of the target DNA initiates LAMP reaction, which proceeds at a constant temperature, followed by strand displacement DNA synthesis primed by an outer primer set. [Futuka et al., 2004, Tomita et al., 2008] [4-5] The addition of two “loop” primers or two “stem” primers further increases the speed of