User:Cassandra M Barrett/Notebook/Open Chromatin/2015/10/02
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Preparation of inserts for LCR
Purpose: to amplify ATF2 and Gal4DB-mCh parts for insertion into MV10. I am repeating this PCR to get higher concentrations of inserts. Phusion didn't amplify well yesterday.
Set up 4 reactions (ATF2, Gal4DB_mCh, and negative controls)
-Template for ATF2: ATF2 mp -FP for ATF2: ATF2 F1 -RP for ATF2: ATF2 R2
-Template for Gal4DB_mCh: KAH228 mp -FP for Gal4DB_mCh: Gal4DB F2 -RP for Gal4DB_mCh: mCh R1
PCRs were run on standard GoTaq protocol (annealing temp: 52C, elongation time: 1min)
Gel shows proper band sizes, both extracted and purified. Good yields!