Standardization of Protocol for Isolation of Genomic DNA from Mature Leaves of Vigna radiata (L.) Wilczek)
2015
Anusheela, V. | Ganesamurthy, K. | Ganeshram, S.
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 mature leaves of Green gram is difficult due to presence of contaminants such as phenols and polysaccharides. Therefore, the present study was under taken to obtain high quality and pure DNA in mature leaves of Green gram. With few modifications three different DNA extraction protocols were tried in the present study to obtain high quality and pure DNA viz., (i) Doyle and Doyle (1987), (ii) Modified Method of Murray and Thompson (1980), and (iii) using of spin column filter. Out of the three methods tried for DNA extraction, the method of Murray and Thompson (1980) 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 Co (Gg) 7.
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