What is the possibility that bacteria doesnt grow? - (Jan/16/2007 )
i did gel extraction after PCR but after that no band is seen after the extraction. i've ligate my PCR product with obtain from Gel Extraction (GE) into the vector and transformed it into the competent cell. i grow it at 37 celcius for 16 hours. no growth is observed. i unable to proceed to pick single colony and do the plasmid extraction. wat do u think it is the possibility and how to overcome it? i've try to read the OD and there are something in there. but i dono whether the thing is it that i want.
Some competent cells needs at least 18h to grow, but there is a possibility that you don't have transformed cells.
If the PCR gives a band only donot make gel extraction (to much DNA lost and could loose also the polyA's), make the ligation directly. If using a kit let the transformation step incubate for a little bit longer than the 5min.