I just checked my plates this morning and absolutely zero colonies grew. I was wondering if anyone has any ideas as to what could be the problem. Below are some things that may have gone wrong. I checked the digested vector and insert and they are in fact the right size. I'm led to believe that the problem is with ligation/transformation
-I used a Qiagen gel extraction kit to get my vector. When I measured the od, there was a high peak at 230 for salt levels. Am I able to just get rid of this with a PCR purification kit?
-I did a 20uL ligation reaction. 15 minutes at room temp. I used 5 uL of that with 50 uL competent cells. 15 minutes in ice. 30 seconds in 42 degree incubator. 2 minutes back in ice. Added 50 uL LB. Warmed up plates for about 30 minutes prior to plating using glass beads.
The competent cells that I have used have been tested and have high efficiency. Vector size is 6000 and insert size is 1000 and I used 3 fold molar excess for ligation.
Any help is much appreciated, thanks in advance.