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- Gibson re-transformation: grow cultures
- Re-transformation: 1x Gibson for pSB1A3 vector assemblies (3 & 4)
Gibson Re-transformation Results (1/06/13)
- Number of colonies did not improve. Only 1 colony on hPCD + BL01 + V0120 plate
- Streaks from yesterday: streak from plate 3 (hPCD + BL01 + pSB1A3 1xGibson) showed no color, whereas streak from plate 7 (hPCD + BL01 + pSB1A3 1/4x Gibson), showed RFP expression. The pSB1A3 template carries RFP.
- Grow 2x 2 mL cultures from plate 3 streak to investigate whether an insert replaced RFP.
- hPCD + BL01 + pSB1A3 (#3), 1xGibson, 6.0 μL
- pSB1A3 (#4), 1xGibson, 6.0 μL
- Add to 50 μL cells; ice/ 10 min.
- Plate on 100 μg/mL amp LB agar