---

=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
     

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

1. 1. 1. [Team:Lethbridge/Notebook|Transformation]]
   • Transformed lumazine synthase - dt in pSB1C3 into DH5∝
   • Transformed pBAD-RBS in pSB1C3 into DH5∝

Contents 1 June 24th, 2011 2 June 25th, 2011 3 June 28th 4 ADD GEL PICTURE 4.1 July 8th

June 24th, 2011

Picked cells from June 21st transformation

June 25th, 2011

Minipreped Cells picked on June 25th, 2011 < br />

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 RFP and lumazine