suman suman

 Perform PCR to amplify DNA sequence encoding GFP from the vector pE-GFP using suitable primers.  Agarose gel electrophoresis for PCR product verification was conducted.  Purification & ligation of GFP DNA into a pQE30 vector using BamHI and HindIII RE  Purification of the digested GFP DNA insert and the cut PQE30 vector using promega SV mini-columns  Transformed E. coli cells with ligated vector Page 3 of 4  Synthesis of GFP by lactose analogue isopropyl ß-D-thiogalactopyranoside (IPTG).  Hexa-histidine tag expressed on protein facilitates its subsequent purification  Hexahistidine-tagged GFP protein was be purified using IMAC (immobilised metal affinity chromatography) on nickel agarose.  Samples was analysed by separating the proteins they contain using SDS-PAGE