Starting with 4 plates:
- HA1 neg control (v0120 E/S digested, no insert)
- EM7:ZeoR neg control (v0120 X/S digested, no insert)
- 20μL in each PCR Tube (16 tubes total): 10μL of gotaq master mix, 8μL of water, 1μL each of the forward and reverse primers
- Use pipette tips to pick 8 colonies from the HA1_v0120 plate and the EM7:ZeoR_v0120 plate, place in PCR tubes for 10 minutes
- Run PCR
- Run the result on a gel to verify that the part is in the vector
Streak one plate with the 8 colonies from the HA1_v0120 plate and another plate with the 8 colonies from the EM7:ZeoR_v0120 plate.
Start Liquid Cultures
Liquid cultures were started from the streaked plates, using HA1_v0120 streaks 1-2 and EM7:ZeoR_v0120 streaks 1-4.