Template:IONIS Paris/Notebook/20-08-15

Results of transformation with the Gibson assembly products

Red colonies there is still some RFP into the tube with purified/digested pSB1C3
We have to try again

Digestion of pSB1C3

Mix digestion preparation:

Tube	pSB1C3
Water	1,5 µL
Buffer 2.1	5 µL
Plasmid	41,5 µL
Enzyme SpeI	1 µL
Enzyme XbaI	1 µL

37°C, 120 min --> heat kill: 80°C, 20min

Dephosphorylation of pSB1C3

Mix plasmid dephosphorylation preparation:

Tube	pSB1C3
Cutsmart	4 µL
Digested Plasmid	25 µL
rSAP	2 µL

37°C, 30 min heat kill: 65°C, 5min

Expected results

Results

Incomplete digestion
Excision of 2000bp bands for gel extraction (QIAgen kit)

20 August 15