DEVELOPMENT OF DNA MARKER USING RESTRICTION DIGESTION ACTIVITY ON VARIOUS PLASMID VECTORS

This study is to report a simple method to prepare high molecular weight DNA ladders. Bacterial culture of DH5-alpha is transformed with pUC18, pYES2/NT A, pYES2/NT C and pYES2/CT vectors. The constructed plasmids are digested using restriction endonucleases to produce specific weights of the DNA fragments followed by purification and precipitation with ethanol. The obtained fragments are mixed proportionally as per the DNA construct, further the obtained fragments are observed on the agarose gel electrophoresis after restriction digestion. These fragments can be used for further studies or can be re-ligated. The DNA fragments obtained are re-ligated into specific plasmid and are transformed into bacteria which can be used for further ligation studies.