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Help for pGEM-T easy vector ligation problem - (Mar/11/2013 )

Hi all,

I have a trouble with ligation of my insert in pGEM-T easy vector. After transformation, I got blue and white colonies. so i took white colonies and did enzyme digestion but there is no insert..i tried ligation for 1hr at RT, overnight at Rt and overnight at 4C..after that no difference, there is only vector..

Please help me...

Thanks in advance



make sure your Polymerase have terminal transferase activity.....otherwise you just gotto optimize the vector and insert ratio....sometimes purifying the PCR product helps....


Thank you so much for your reply....