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Work to be done:

Contents 1 Week of June 14/2010 1.1 Test T4 DNA Ligase: 1.2 Test PCR conditions for confirmation of ligation via PCR: 1.3 PCR Confirm previous ligations

Week of June 14/2010

Test T4 DNA Ligase:

• Restrict rbs-xylE with EcoRI and SpeI (take sample for analysis on gel)
  
• Heat kill EcoRI and SpeI.
• Ligate with T4 DNA Ligase
• Run unrestricted, restricted and ligated samples on a gel.

Test PCR conditions for confirmation of ligation via PCR:

• Run a PCR of lumazine synthase gene that was sent for sequencing (and subsequently confirmed)
  • using VF2 and VR primers
• Run this PCR on a 2% agarose gel

PCR Confirm previous ligations

If (AND ONLY IF) PCR conditions amplified our known lumazine synthase gene, set up PCR of our previous ligation reactions to see if there is any ligated product.