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no colonies - Cloning using pENTR vector, no colonies (Mar/25/2009 )

I' m trying to do cloning using pENTR vector, but after transformation i got no colonies in my plates (LB+ kan+Hyg)
i need help!


Leave out the Hyg


I am new to cloning also but I haven't had any problem with not having colonies on the agar so far. but I have this colleague who couldn't get colonies on agar although we did all the steps together, then we realized she used too much kanamycin in the agar. now it is working fine for her.

use 50ug/ml. we do not use Hyg.