Standardization of DNA Extraction Protocol in Greengram (Vigna radiata (L.) Wilczek).

ABSTRACT

Green gram is a widely cultivated pulse crop rich in protein, high in vitamin-B content and essential aminoacids. It is easily digestable and low flatulence produced crop. The quality and quantity of DNA used for amplification by PCR is the key to reproducible results and success of genotyping. Especially, DNA purity is extremely crucial for obtaining clear and discriminate patterns. DNA extraction from Green gram is difficult due to presence of contaminants such as phenols. Therefore, the present study was under taken to obtain high quality and pure DNA in Green gram. With few modifications four different DNA extraction protocols were tried in the present study to obtain high quality and pure DNA viz., (i) Doyle and Doyle (1987), (ii) Method of Murray and Thompson (1980), (iii) Porebski et al.(1997), and (iv) Lin et al. (2001). Out of the four methods tried for DNA extraction, the method of Lin et al. (2001) was found most efficient, as the DNA obtained through this protocol was relatively pure which gave amplifying products in the PCR. The genotype used for the standardization was MGG -361.