Team:Hawaii/Notebook/2008-08- 6

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(New page: {{Team:Hawaii/Header}} = Things we did today = == Wetlab work == ===Checked transformants from yesterday=== :<strong> Grace</strong> :* Colony PCR'd transformants ::* 30 cycles, anneal a...)
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:<strong> Grace</strong>
:<strong> Grace</strong>
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{|class=wikitable border=1 align=center
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!Construct
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!Colony forming units
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|-
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|align=center|I14032 (plac) + B0030 (rbs)
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| align=center|3
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|-
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|align=center|pnir + B003 (rbs)
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|align=center| 0
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|-
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|align=center|E0040 (GFP) + B0015 (tt)
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|align=center| 1
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|-
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|align=center|GFPf + B0015 (tt)
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| align=center|2 + 2 clusters of colonies
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|}
:* Colony PCR'd transformants
:* Colony PCR'd transformants
::* 30 cycles, anneal at 62C, extend for 1 min.
::* 30 cycles, anneal at 62C, extend for 1 min.

Revision as of 02:13, 7 August 2008

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Notebook (t) Meetings (t)

Things we did today

Wetlab work

Checked transformants from yesterday

Grace
Construct Colony forming units
I14032 (plac) + B0030 (rbs) 3
pnir + B003 (rbs) 0
E0040 (GFP) + B0015 (tt) 1
GFPf + B0015 (tt) 2 + 2 clusters of colonies
  • Colony PCR'd transformants
  • 30 cycles, anneal at 62C, extend for 1 min.
  • Ran on EtBr stained 2.0% agarose gel at 95V for 1 hour
  • None of the transformations were successful :o(

Prepared PCR'd nir and J33207 (from yesterday) for transformation/ligation

Grace
  • Ran PCR products on EtBr stained 2.0% agarose gel at 60V for 100 min.
  • nir band at 330bp confirmed
  • J33207 = 4 bands (1.5kb, 2.5kb, 3.2kb, 10kb), none of which are correct
  • [http://www.partsregistry.org partsregistry] says J33207 DNA is inconsistent
  • Extracted nir and 1.5kb J33207 band from gel
  • RE digested in 50 μl rxns with EcoRI and SpeI in NEBuffer 2
  • Larger rxn volume may improve digest efficiency
  • Incubated at 37C for 2.5 hours
  • RE digested 10 μl J33207 plasmid prep with EcoRI and SpeI in NEBuffer 2
  • To confirm plasmid prep; if good, will use for ligation rxn
  • Incubated at 37C for 2 hours
  • Ran RE digests on EtBr stained 2.0% agarose gel at 72V for 100 min.

Made LB+amp100 plates

Grace and Margaret

Drylab work

Sequencing

Grace
  • Reply from CORE Hawaii

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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