June

June 21st, 2011

Restriction, Ligation, and Transformation Lumazine synthase - double terminator and pBAD - RBS into pSB1C3

1. Restriction

• Restricted lumazine synthase - dt out of pSB1K3 and pBAD - RBS out of pSB1T3 with EcoR1 and Pst1
• Restricted destination plasmid pSB1C3 with EcoR1 and Pst1

2. Ligation

• Ligated lumazine synthase-dt into destination plasmid pSB1C3
• Ligated pBAD-RBS into destination plasmid pSB1C3

3. Transformation

• Transformed lumazine synthase - dt in pSB1C3 into DH5∝
• Transformed pBAD-RBS in pSB1C3 into DH5∝

June 24th, 2011

Picked cells from June 21st transformation

June 25th, 2011

Minipreped Cells picked on June 25th, 2011

June 28th

Restriction digest and ran on gel
Lumazine - dt construct successfully inserted into pSB1C3 Glycerol stocked.

Add Gel Picture

July 8th

Performed antibiotic assay on lumazine - dt in pSB1C3 Samples from one colony grew in Kanamicyn; this stock was discarded

PCR PCR'd out lumazine - dt