Difference between revisions of "Team:CSU Fort Collins/NoteBook/JanThroughApr"

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<li>Add 1uL phosphatase and 5uL 10x buffer to bring total rxn volume to 50uL</li>
 
<li>Add 1uL phosphatase and 5uL 10x buffer to bring total rxn volume to 50uL</li>
 
<li>Incubate at 37C for 1 hour, and heat inactivate at 70C for 5 minutes</li>
 
<li>Incubate at 37C for 1 hour, and heat inactivate at 70C for 5 minutes</li>
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<div class = 'Days'><h5>1/25/16:</h5></div>
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<div class = 'Info'><ul><li>Overnight ligated R0062 and I0462, B0034 and C0061, B0015 and E0030 at 16 C</li>
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</ul>
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<div class = 'Days'><h5>1/25/16:</h5></div>
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<div class = 'Info'><ul><li>Transformed the overnight ligation mixtures</li>
 
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</ul>
 
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<div class = 'Info'><ul><li>To confirm that Bba_K325909 are functioning we tried to induce expression using L-arabinose</li>
 
<div class = 'Info'><ul><li>To confirm that Bba_K325909 are functioning we tried to induce expression using L-arabinose</li>
 
<li>No luminescence was produced</li>
 
<li>No luminescence was produced</li>
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</ul>
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</div>
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<div class = 'Days'><h5>2/16/16:</h5></div>
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<div class = 'Info'><ul><li>Decided to order two gBlocks for quorum portion of the project</li>
 
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</ul>
 
</ul>
 
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<div class = 'Days'><h5>4/12/16:</h5></div>
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<div class = 'Info'><h6>Prepare pSB1C3 vector for Infusion</h6><ul><li>gBlocks (Quorum 1 and Quorum 2) have 15 bp homology with each other along with prefix and suffix at the entire block’s ends</li>
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<li>Digesting the vector with XbaI will leave 16 bp homology with Quorum 1</li>
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<li>Digesting the vector with SpeI will leave 20 bp homology with Quorum 2</li>
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<li>Digested c0061 with SPeI and XbaI</li>
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<li>Ran on 1% gel</li>
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<img src = "https://static.igem.org/mediawiki/2016/a/a8/CSU_g41216.png">
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Revision as of 22:39, 16 October 2016

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CSU iGEM



January Through April

1/4/16:
  • Ligated b0034 and c0061
  • Transformed Ligated mixture
  • Ran PCR results on a 2% gel
1/5/16:
  • Digested LuxR with EcoRI and SpeI
  • Performed Colony PCR on transformation from the 4th
1/11/16:
  • slr0435 Excision from Synechocystis sp. PCC6803 with PCR using Phusion(size 276 bp).
  • Ran PCR results on a 2% gel
  • Digested I0462 and E0030 with EcoRI and SpeI, R0062 and B0015 with EcoRI and XbaI, B0034 with SPeI and PstI, and C0061 with XbaI and PstI.
1/12/16:
  • Digested slr0435 and resuspended pSB1C3 with EcoRI and PstI
  • Ligated pSB1C3 and slr0435
  • Transformed ligation mixture
1/13/16:
  • Colony PCR performed on slr0435+pSB1C3 transformed colonies
  • Ran PCR results on 1% gel
  • Digested I0462 and E0030 with EcoRI and SpeI, R0062 and B0015 with EcoRI and XbaI, B0034 with SPeI and PstI, and C0061 with XbaI and PstI.
  • Ran on 1% gel
1/21/16:
  • Digested I0462 and E0030 with EcoRI and SpeI, and C0061 with XbaI and PstI.
  • Ran on 1% gel
  • Phosphatase treatment
  • Use Antarctic Phosphatase with 10x buffer. Need 1uL to treat 1ug of DNA
  • Add 1uL phosphatase and 5uL 10x buffer to bring total rxn volume to 50uL
  • Incubate at 37C for 1 hour, and heat inactivate at 70C for 5 minutes
1/25/16:
  • Overnight ligated R0062 and I0462, B0034 and C0061, B0015 and E0030 at 16 C
1/25/16:
  • Transformed the overnight ligation mixtures
2/6/16:
  • PCRed out BBA_J23106 out of glycerol stock 19 using Q5 master mix 19rxns
2/9/16:
  • Transformed Bba_K325909 from parts kit into DH5a cels
2/11/16:
  • To confirm that Bba_K325909 are functioning we tried to induce expression using L-arabinose
  • No luminescence was produced
2/16/16:
  • Decided to order two gBlocks for quorum portion of the project
3/24/16:
  • Set up O/N cultures of K1033929, K592009 and E1010 in 5 mL of LB with 5 uL of Cm34 antibiotic
3/29/16:
  • Ran promoters Sll027, and LrtA in a 1% gel
    • Sll0027
    LrtA
4/6/16:
  • Ran Cpcg2 on 1% gel
4/12/16:
Prepare pSB1C3 vector for Infusion
  • gBlocks (Quorum 1 and Quorum 2) have 15 bp homology with each other along with prefix and suffix at the entire block’s ends
  • Digesting the vector with XbaI will leave 16 bp homology with Quorum 1
  • Digesting the vector with SpeI will leave 20 bp homology with Quorum 2
  • Digested c0061 with SPeI and XbaI
  • Ran on 1% gel