APPLICATION AND COST-EFFECTIVENESS OF LAMP (LOOP-MEDIATED ISOTHERMAL AMPLIFICATION) TECHNOLOGY FOR AUTHENTICATION OF MEDICINAL PLANTS PODOPHYLLUM HEXANDRUM ROYLE

摘要

We have developed a novel LAMP-based molecular marker for easy and fast identification of Podophyllum hexandrum Royle (Indian May apple). It is an important medicinal plant valued all over the world. It have both anti-cancer (pro-oxidant ability) and radioprotective (antioxidant ability) effects respectively. The LAMP-based molecular was constructed based on genomic DNA of P. hexandrum that amplified by RAPD. A RAPD amplicon common to all accessions, was eluted, cloned and sequenced. Based on these sequences in-house specific LAMP primers were synthesized. This method employs by a Bst DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the P. hexandrum genomic DNA. In LAMP reaction the amplification was carried out in thermal cycler and water bath at 60°C for 60 min, which was followed by monitoring DNA amplification with the naked eye through color changes by standardizing SyBr Green? I nucleic acid stain. The data showed confirmatory results in both thermal cycler and water bath. Since, the assay method is sensitive and cost-effective; it is a feasible method for identifying and authentication of P. hexandrum. Due to its specificity and simplicity, the method could become a useful tool for identification and authentication medicinal plants.