Cultures for a few of the constructs didn't grow:
hEF1a:Gal4
hEF1a:CD79B-TCS-G4
EGSH:mKate
hEF1a:VgEcR-2A-RxR
hEF1a:CymR | hEF1a-CuO:mKate | TRE:Syk | EGSH:mKate | hEf1a:VgEcR-2A-RXR | TRE:mKate | hEF1a:Gal4VP16 | hEF1a:CD79B-TCS-Gal4VP16 | hEF1a:tagBFP | |
---|---|---|---|---|---|---|---|---|---|
Enzyme | PstI Cuts in hEF1a, CymR and backbone | StuI Cuts in hEF1a, mKate and backbone | StuI, ApaI Cuts in TRE, Syk and backbone | ApaI Cuts in hEF1a, VgEcR-2A-RxR and bacbone | StuI cuts in TRE, mKate, and backbone | XhoI cuts in hEF1a, Gal4VP16, and backbone | PvuII Cuts in CD79B, TCS and backbone | StuI Cuts in hEF1a, tagBFP and backbone | |
digest tube | 4 | 5 | 6,7,8 | 1 | 2,3 | ||||
4 | 5 | 6,7,8 | 1 | 2,3 | |||||
Virtual gel |
Ladder, TRE:mkate, tagBFP 1, tagBFP2, hEF1a:CymR, CuO:mKate, TRE:syk(double digest), TRE:syk(StuI), TRE:Syk(undigested
| Notes: TRE:mKate - extra band at 2.5? tagBFP - both correct hEF1a:CymR - correct CuO:mKate - correct TRE:Syk - correct |
Picked new colonies for cultures that didn't grow + TRE:mKate