I have a problem with my yeast colony PCR.
I am including my yeast colony PCR protocol here:
1. Incubate yeast culture with medium at 30°C, overnight.
2. Then centrifuge 0.5ml of cells at 8000rpm for 5mins, after discard supernatant.
3. Resuspend the cells with 0.5ml 1xPBS, centrifuge at 8000rpm for 5mins,discard supernatant.
4. Resuspend the cells with 0.1ml TE, then boil in water for 3 mins
5. cold in the liquid nitrogen for 0.5min
6. boiled in water for 2mins.
7.then take 1ul supernatant as template for PCR
The problem is that i cannot get the right PCR product, I actually get nothing.
Is this yeast colony protocol ok?