User:Mara Peterson/Notebook/Mara Peterson/2015/12/15

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Starting with 4 plates:

  • HA1_v0120
  • EM7:ZeoR_v0120
  • HA1 neg control (v0120 E/S digested, no insert)
  • EM7:ZeoR neg control (v0120 X/S digested, no insert)

Colony PCR

  • 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 Plates
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.