Abstract

DEVELOPMENT OF LOOP MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) MARKER FOR AUTHENTICATION FOR VARIETY OF STEVIA REBAUDIANA (BERTONI)

Sunanda Singh*, Veena Garg and Deepak Yadav

ABSTRACT

A LAMP-based molecular marker was designed for simple and rapid identification of medicinal plant Stevia rebaudiana (Bertoni) for five different accessions over India. The LAMP-based molecular was developed through randomly amplified polymorphic DNA (RAPD). A common RAPD band among all accessions was eluted and cloned. Now it was sequenced and LAMP primers were designed by these sequences. This technique hires a DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA. In LAMP reaction the amplification was carried out in a water bath at 65°C for 50 min, which was followed by monitoring DNA amplification with the naked eye through color changes with the help of SYBER Green I. The data showed confirmatory florescence under UV transilluminator. Hence, the technique is quick, delicate and reasonable; it is a feasible method for identifying and authentication of S. rebaudiana. The method could become a beneficial and convenient technique for medicinal plant S. rebaudiana so that adulterants can identified easily.

Keywords: Loop-Mediated Isothermal Amplification (LAMP), Stevia rebaudiana, Authentication, Polymerase Chain Reaction (PCR), RAPD.