I have a problem with cloning pHannibal cassette in pART27....I cut phannibal cassette and open pART27 with NotI and seems to be OK....after i treat the pART27 with CIAP, extract all from gel...ligate and cloning the ligation in E.coli DH5alfa...after i select on antibiotic Kanamicin 50ug/ml...but not works....There's anyone that can help me????

Vinny:

In the past, I have had problems with my antibiotic concentration being too high during selection. You may want to try and select at a lower concentrations such as 25 ug/ml.