Team:Hawaii/Notebook/2008-09-11

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(Difference between revisions)
(New page: {{Team:Hawaii/Header}} = Things we did today = == Wetlab work == ===Construction of p+r + s(+g<sub>f</sub>)=== :<strong> Grace</strong> [[Image:091108REdigests.jpg|right|thumb|200px|EtBr ...)
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::* plac+rbs (on SpeI/PstI digested pSB1A2 plasmid) with slr1, pilA, slr1+GFPf, pilA+GFPf (8/12 and 8/13 transformations)
::* plac+rbs (on SpeI/PstI digested pSB1A2 plasmid) with slr1, pilA, slr1+GFPf, pilA+GFPf (8/12 and 8/13 transformations)
:* Transformed into DH5&alpha; (3A ligations) or DB3.1 (2-way ligations) cells
:* Transformed into DH5&alpha; (3A ligations) or DB3.1 (2-way ligations) cells
 +
===Restriction digest===
 +
:<strong>Krystle</strong>
 +
::20ul reactions:
 +
:*RE digest 3 versions of GFPf+tt with XbaI + PstI
 +
:*RE digest rbs+GFP with EcoRI + SpeI
 +
 +
===Plasmid Prep====
 +
:<strong>Krystle</strong>
 +
:* 5ml plasmid preps of nir, pilA, slr1 and slr2
= Discussion =
= Discussion =

Revision as of 03:59, 12 September 2008

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Things we did today

Wetlab work

Construction of p+r + s(+gf)

Grace
EtBr stained 4% agarose gel ran at 47V for 2.5 hours. Thirty microliters of RE digest reaction were loaded into each well.
  • Ran RE digests from last night on gel
  • Extracted DNA from gel
  • slr1, pilA, nir+rbs
  • (From yesterday) slr1+GFPf, pilA+GFPf (8/12 and 8/13 transformations)
  • Ligated:
  • pSB1A3 and nir+rbs with slr1, pilA, slr1+GFPf, pilA+GFPf (8/12 and 8/13 transformations)
  • nir+rbs (on SpeI/PstI digested pSB1A2 plasmid) with slr1, pilA, slr1+GFPf, pilA+GFPf (8/12 and 8/13 transformations)
  • plac+rbs (on SpeI/PstI digested pSB1A2 plasmid) with slr1, pilA, slr1+GFPf, pilA+GFPf (8/12 and 8/13 transformations)
  • Transformed into DH5α (3A ligations) or DB3.1 (2-way ligations) cells

Restriction digest

Krystle
20ul reactions:
  • RE digest 3 versions of GFPf+tt with XbaI + PstI
  • RE digest rbs+GFP with EcoRI + SpeI

Plasmid Prep=

Krystle
  • 5ml plasmid preps of nir, pilA, slr1 and slr2

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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