A Simple and Efficient Method For Extraction of Genomic Dna from Tropical Tuber Crops |

ABSTRACT:

DNA extraction in many plants is difficult because of metabolites thatinterfere with DNA isolation procedures and subsequent applications, such asDNA restriction, amplification and cloning. We have developed a reliable andefficient method for isolating genomic DNA free from polysaccharide,polyphenols and protein contaminants from tropical tuber crops (Elephant footyam, Cassava, Sweet potato Taro and Tannia). The method involves inactivationof contaminant proteins by using CTAB/Proteinase K and precipitation ofpolysaccharides in the presence of high concentration of salt. The purity of genomic DNA was confirmed by A260/280 and A260/230ratios calculated from the spectrophotometric readings and further byrestriction analysis of the isolated DNA using restriction enzymes EcoRIand Hind III. The described protocol also resulted in the isolation ofsufficiently higher yield of DNA from leaf sample of tropical tuber crops. Thenew protocol can be successfully used for both small and large scalepreparation of genomic DNA from leaf tissues of tuber crops which is highlysuitable for further down stream processes like PCR amplification andrestriction digestion analysis.