PCR with cell source - (Oct/13/2001 )
Does anyone know of a protocol for PCR where I can start with whole cells and lyse them right there in the PCR tubes as I'm doing it?
I use this method routinely for colony screening. If you are using bacterial cells.... Make up your PCR cocktail as normal and aliquot into as many tubes as needed. With a toothpick or tip pick one bacterial colony and swirl tip into cocktail. Don't worry you will have enough DNA. As the temperature raises to 95 degrees the cells will lyse. You can also save the tip in a culture tube during the PCR, that way when you know which are your positive clones you can start O/N cultures for minipreps.