Sunday 2^nd October

Lab work

Visualization

Cloning of FRB - GFP 11 (pPS16_019) in FKBP - GFP 10 - pSB1C3 (pPS16_018) by digestion-ligation

By Maxence & Victor

PRECIPITATION ETOH

Once template and vector were cut, DNA ligase was used to join the sticky ends of the template and vector together:

• 1.5 µL of Buffer T4 10X
• 1 µL of ligase T4 enzyme
• 1 µL of ligase T4 enzyme
• 12.5 µL of water

The mix were incubated for 30 minutes at rooming temperature.

Cloning of GFP 1.9 from pUC19 (pPS16_009) in pSB1C3 by digestion-ligation

By Maxence & Victor

Once template and vector were cut, DNA ligase was used to join the sticky ends of the template and vector together:

• 6 µL of template (pPS16_009 treated by PstI & XbaI)
• 3 µL of vector (Bba treated by PstI & XbaI)
• 1.5 µL of Buffer T4 10X
• 1 µL of ligase T4 enzyme
• 3.5 µL of water

The mix were incubated for 30 minutes at rooming temperature.

Plasmids extraction of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6 and GFP 1.9 in pUC19 (pPS16_009) clone 1

"By Maxence & Victor"

The following plasmids were extracted using the "Charge Switch-Pro Plasmid Miniprep" kit:

• pPS16_018 (FKBP - GFP 10) clone 6
• pPS16_019 (FRB - GFP 11) clone 4
• pPS16_009 (GFP 1.9) clone 1