Team:Hawaii/Notebook/2008-07-14

From 2008.igem.org

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:* Incubated on ice 10 min as opposed to the recommended 30 min. by iGEM
:* Incubated on ice 10 min as opposed to the recommended 30 min. by iGEM
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===Transformed DH5α with mutated GFP and new Biobrick vector constructs===
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===RE digested PCR products and plasmids===
:<strong> Grace</strong>
:<strong> Grace</strong>
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:* Constructs:
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:* XbaI and PstI: GFP fusion brick and BBa_C0012  
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::* PCR'd GFP fusion brick ligated with Biobrick vector derived from BBa_C0012
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::* Added XbaI and PstI separately
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::* Biobrick pertinent segment ligated with pRL1383a
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:* HindIII and BamHI: Biobrick segment and pRL1383a  
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::* Constructs were restriction digested, agarose gel purified, and ligated together using Quick Ligase
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::* pRL1383a was not visbile under long wave UV. Need to rerun gel tomorrow with much more plasmid
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===Transformed DB3.1===
===Transformed DB3.1===

Revision as of 04:18, 15 July 2008

Projects Events Resources
Sponsors Experiments Milestones Protocols
Notebook (t) Meetings (t)

Things we did today

Wetlab work

PCR

Grace
  • 25 μl PCR reactions for GFP site directed mutagenesis and extraction of Biobrick pertinent sites from BBa_B0034

Made stocks

Grace
  • Made two 500mL batches of SOB
  • Made 20 LB + amp100 plates

Transformed DH5α with ligations of a Biobrick vector with pnir, slr2016-1, slr2016-2, pilA, or C0012

Grace
  • Transformed using 5 μl of 1:1 and 1:10 ligation dilutions
  • Transformed using 1 μl vector only (negative control)
  • Incubated on ice 10 min as opposed to the recommended 30 min. by iGEM

RE digested PCR products and plasmids

Grace
  • XbaI and PstI: GFP fusion brick and BBa_C0012
  • Added XbaI and PstI separately
  • HindIII and BamHI: Biobrick segment and pRL1383a
  • pRL1383a was not visbile under long wave UV. Need to rerun gel tomorrow with much more plasmid

Transformed DB3.1

Margaret
  • Transformed: pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-)nothing

Discussion

Quote of the Day

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson


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