1. Transfer 5 ml overnight culture
to a microcentrifuge tube or 96 well plate containing 95
ml of ddH2O.
For clones on a LB plate, pick single clones using toothpicks, and
resuspend cells in 100 ml of ddH2O.
2. Boil for 5 minutes or 95 °C for 10 minutes if using 96 well plate to lysis cells.
3. Remove cell debris by brief centrifugation. For 96 well plate, centrifuge at 1200xg for 3 minutes.
4. Clones are amplified in 50 ml PCR reactions. Make a master
mix, add 48 ml of master mix to 2 ml of DNA.
5. Run 5 ml on an argarose gel to check for amplifications.