Team:Hawaii/Notebook/2008-07-14

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(New page: {{Team:Hawaii/Header}} = Things we did today = == Wetlab work == ===PCR=== :<strong> Grace</strong> :* 25 μl PCR reactions for GFP site directed mutagenesis and extraction of Biobrick...)
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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 DH5&alpha; 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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::* Ran 25 &mu;l of GFP and 20 &mu;l of BBa_C0012 restriction digest reactions in gel
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::* Constructs were restriction digested, agarose gel purified, and ligated together using Quick Ligase
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:* HindIII and BamHI: Biobrick segment and pRL1383a  
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::* Ran 25 &mu;l of Biobrick segment (half of total rxn) and 20 &mu;l of pRL1383a restriction digest reactions in gel
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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===
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:<strong> Margaret</strong>
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:*Transformed: pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-)nothing. The positive and negative control verify that the DB3.1 competent cells  made on 7/11/08 can take up DNA. The results of this test can be seen [[Team:Hawaii/Test Competent E. Coli|here]].
= Discussion =
= Discussion =
= Quote of the Day =
= Quote of the Day =
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<blockquote>''History is the only laboratory we have in which to test the consequences of thought.'' - Étienne Gilson</blockquote>
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<blockquote></blockquote>
{{Team:Hawaii/Footer}}
{{Team:Hawaii/Footer}}
__NOTOC__
__NOTOC__

Latest revision as of 01:23, 16 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
  • Ran 25 μl of GFP and 20 μl of BBa_C0012 restriction digest reactions in gel
  • HindIII and BamHI: Biobrick segment and pRL1383a
  • Ran 25 μl of Biobrick segment (half of total rxn) and 20 μl of pRL1383a restriction digest reactions in gel
  • 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. The positive and negative control verify that the DB3.1 competent cells made on 7/11/08 can take up DNA. The results of this test can be seen here.

Discussion

Quote of the Day


[http://manoa.hawaii.edu/ Sponsor_UHM.gif][http://manoa.hawaii.edu/ovcrge/ Sponsor_OVCRGE.gif][http://www.ctahr.hawaii.edu Sponsor_CTAHR.gif]


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