Team:Hawaii/Notebook/2008-08-16

From 2008.igem.org

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(Reconstruction of BB-pRL1383a (cont.))
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= 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>''She grew on him like she was a colony of E. coli, and he was room temperature Canadian beef.'' - Courtesy of [http://help.com/post/124066-funny-metaphors-used-in-high-school Help.com]</blockquote>
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Revision as of 01:05, 17 August 2008

Projects Events Resources
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Notebook (t) Meetings (t)

Things we did today

Wetlab work

Reconstruction of BB-pRL1383a (cont.)

Grace
EtBr stained 1% agarose gel ran at 60V for 2 hours. Thirty microliters of RE digested DNA were loaded into each well.
  • Ran digests on a 1% agarose gel
  • J33207 bands @ ~900bp and smudge ~100bp. ~100bp=VF/VR? Too much DNA loaded = band ran slower?
  • BB-pRL1383a band @ ~10kb. No band ~750bp. XbaI/PstI did not cut??
  • Extracted bands from gel
  • Ligated:
  • 1 μl XbaI/PstI digested BB-pRL1383a with 3 μl XbaI/PstI digested J33207
  • 1 μl XbaI/PstI digested BB-pRL1383a to itself (negative control)
  • Transformed into DH5α
  • Used 5 μl ligation reaction to transform
  • Plated on LB+sp100+X-gal and LB+sp100+sm50+IPTG+X-gal
  • To verify if IPTG is needed for lacZ expression

Discussion

Quote of the Day

She grew on him like she was a colony of E. coli, and he was room temperature Canadian beef. - Courtesy of [http://help.com/post/124066-funny-metaphors-used-in-high-school Help.com]


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