Team:CSU Fort Collins/NoteBook/July

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CSU iGEM



July

7/6/16:
  • PCRed out Lrta from Synechocystis PCC 6003
7/7/16:
  • Ran Lrta PCR products on a 1% gel
7/8/16:
  • Performed ligase cycling reaction on slr0168 and pSB1C3
  • Ran all PCR products on 1%gel
  • Digested Lrta(SpeI EcorI)
  • Ligated Lrta and EYFP-LVA
  • Transformed Lrta+EYFP-LVA ligation mixture
7/12/16:
  • Dpn1 Digested slr0168 and pSB1C3
  • Phosphorylated slr0168 and pSB1C3
  • Performed ligase cycling reaction on phosphorylated mixture
  • Transformed ligase cycling reaction products
  • Digested EYFP-LVA(PstI)
  • Ligated EYFP-LVA + LrtA
  • Transformed EYFP-LVA+LrtA ligation mixture
  • Performed colony PCR on 13 colonies with the Q1 block
  • Ran PCR results on a 1.5% gel
7/13/16:
  • Colony PCR performed on 10 EYFP-LVA+LrtA transformed colonies
  • PCR amplified Q1 and Q2 for infusion cloning
7/14/16:
  • Ran Q1 and Q2 blocks in a 1.5% gel
  • Gel had no bands
  • PCR amplified for infusion cloning
7/15/16:
  • Ran Q1 and Q2 blocks in a 1.5% gel
  • Infusion cloning on Quorum and PSB1C3
  • Transformed infusion mixture
7/16/16:
  • PCRed PSB1C3 for infusion cloning
  • Ran PCR products on a 1.5% gel
  • Infusion performed on Quorum gBlocks and PSB1C3
  • Transformed infusion product
7/19/16:
  • 2-step PCRed Qblocks together
  • Ran PCR products on a 1.5% gel
7/20/16:
  • Redid Qblock 2-step PCR
  • Ran PCR products on a 1.5% gel
7/27/16:
  • Q1,Q2,PSB1C3 were all PCRed for ligase cycling
7/27/16:
  • Ran PCR products on a 1.5% gel
7/29/16:
  • Dpn1 DIGESTED Q1 and Q2 PCR products
7/30/16:
  • Phosphorylated PSB1C3,Q1,and Q2 for ligase cycling
  • Ligase cycling performed on phosphorylated mixture
  • Transformed ligase cycling mixture