cloning into bacterial expression vector - problem in E.coli ttransformation (Jan/03/2009 )
I am trying to subclone pcr product from a plasmid into the bacterial expression vector. ligation product shows the expected Molecular weight. no problem with the competant cells and transformation protocols as the control reaction set up showed positive colonies. no colonies appear on the transformation plate.
It happens. I see it all the time, you just need to repeat it. Make sure that the vector and insert used for ligation is not too much. I have a rule which i follow, If my clong doesn't work once, I start from scratch coz I don't know where have i made a mistake (if I have).