Team:Hawaii/Notebook/2008-08-25

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(Difference between revisions)
(→Verification of transformants (cont.))
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::* C0012 with EcoRI and PstI (to isolate vector)
::* C0012 with EcoRI and PstI (to isolate vector)
::* nir with EcoRI and SpeI
::* nir with EcoRI and SpeI
 +
 +
===Ligation===
 +
:<strong>Krystle</strong>
 +
 +
:* Ligated overnight:
 +
::* plac + B0034 (both synthesized)
 +
::* plac + B0030 (both synthesized)
 +
::* gfpf (vector) + tt (synthesized)
 +
::* gfpf (re'd with E-S) + tt (synthesized)
= Discussion =
= Discussion =

Latest revision as of 03:05, 27 August 2008

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

Things we did today

Wetlab work

Ligation & Transformation

Margaret
  • ligation of rep+pSB1A3 and P1+pSB1A3, transformation into DH5alpha cells

Colony PCR

Margaret

  • verified the colonies from the (pLac+B0030) ligation and transformation. no transformants for the B0034 ligation

Verification of transformants (cont.)

EtBr stained 3% agarose gel ran at 60V for 3 hours. Two microliters of PCR reaction were loaded into each well.
Grace
  • PCR'd nir, nir+rbs, GFP, GFP+rbs
  • Ran on 3% agarose gel

Restriction digest

Grace
  • RE digested overnight:
  • BB-pRL1383a and J33207 with XbaI and PstI
  • C0012 with EcoRI and PstI (to isolate vector)
  • nir with EcoRI and SpeI

Ligation

Krystle
  • Ligated overnight:
  • plac + B0034 (both synthesized)
  • plac + B0030 (both synthesized)
  • gfpf (vector) + tt (synthesized)
  • gfpf (re'd with E-S) + tt (synthesized)

Discussion

Quote of the Day

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


[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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