Team:Hawaii/Notebook/2008-08- 8

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===Restreaked nir+rbs and I14032+rbs constructs===
===Restreaked nir+rbs and I14032+rbs constructs===
:<strong> Grace</strong>
:<strong> Grace</strong>
 +
 +
===Plasmid prep (finished up)===
 +
:<strong>Grace</strong>
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 +
:* Resuspended plasmid preps in 50 &mu;l TE buffer
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:* Determined DNA concentrations of plasmids
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{|class=wikitable border=1 align=center
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!Plasmid
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! DNA Concentration
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|-
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|align=center |nir
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|align=center |496.3 ng/&mu;l
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|-
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|align=center |GFPf
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|align=center |484.8 ng/&mu;l
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|-
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|align=center |BB-pRL1383a
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|align=center |508.0 ng/&mul
 +
|}
===Prep for sequencing===
===Prep for sequencing===

Revision as of 18:52, 8 August 2008

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Notebook (t) Meetings (t)

Things we did today

Wetlab work

Restreaked nir+rbs and I14032+rbs constructs

Grace

Plasmid prep (finished up)

Grace
  • Resuspended plasmid preps in 50 μl TE buffer
  • Determined DNA concentrations of plasmids
Plasmid DNA Concentration
nir 496.3 ng/μl
GFPf 484.8 ng/μl
BB-pRL1383a 508.0 ng/&mul

Prep for sequencing

EtBr stained 4% agarose gel ran at 60V for 100 min. Twenty-five microliters of the PCR reactions were loaded into each well.
Grace
  • 25 μl PCR reactions of nir+rbs, I14032+rbs, slr1, slr2, BB-pRL1383a
  • Colony PCRs seem to indicate success
  • 25 μl PCR reactions of GFP+tt, GFPf+tt, J33207+tt
  • Colony PCR indicates no ligation. Picked new colony, sequence to confirm failure.
  • Gel purified all PCR rxns (we still have a problem with contaminant DNA/shadow bands) and desired bands were extracted from gel
  • 2% agarose gel ran at 60v for 2 hours
  • Determined DNA concentrations via nanodrop spectrometer
  • Prepared samples and sent to CORE Hawaii for sequencing

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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