Team:Washington/Notebook/colony PCR

Colony PCR

 * 1) Prepare one sterile 0.6mL tube with the following reaction mixture for each colony you intend to pick.
 * 2) 5uL Qiagen Master Mix
 * 3) 1uL 40uM VF2
 * 4) 1uL 40uM VR
 * 5) Prepare one sterile 0.6mL tube with 20uL sterile diH2O for each colony you intend to pick.
 * 6) Pick colonies
 * 7) Pick a single colony using a micropipettor with sterile tip. The pippettor should be set to 3uL
 * 8) Aspirate colony into 20uL diH2O vigorously to transfer cells to diH2O
 * 9) Transfer 3uL of diH2O containing cells to reaction mixture set up in step 1
 * 10) Run reaction
 * 11) 94C - 3min
 * 12) 94C - 30s
 * 13) 55C - 30s
 * 14) 72C - 1min / kb gene
 * 15) repeat 2 - 4 29x
 * 16) 72C - 10min
 * 17) 4C - forever