Team:Hawaii/Notebook/2008-09- 5

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(Difference between revisions)
(New page: {{Team:Hawaii/Header}} = Things we did today = == Wetlab work == ===Construction of Broad-Host-Range Vector Intermediates=== :<strong> Margaret</strong> :* Plasmid Prep: Everything that ...)
(→Construction of Broad-Host-Range Vector Intermediates)
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:*Ligation
:*Ligation
:*Transformation into DH5alpha (we need more competent cells)
:*Transformation into DH5alpha (we need more competent cells)
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===Verification of transformants from yesterday===
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:<strong>Grace</strong>
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[[Image:090508colonypcr.jpg|right|thumb|400px|EtBr stained 2% agarose gel ran at 83V for 1.75 hours. Two microliters of PCR product were loaded into each well. ]]
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{|class=wikitable border=1 align=center
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!Construct
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!Colonies
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|-
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|align=center|plac+rbs (no vector)
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|align=center|0
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|-
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|align=center|C0012 derived vector
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|align=center|12
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|-
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|align=center|C0012 derived vector, SAP treated
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|align=center|163
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|-
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|align=center|3A: plac+rbs+vector
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|align=center|>300
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|}
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:* BBpRL1338a related plates all had a lawn (no distinct colonies observed). Too low antibiotic conc.? Will attempt to restreak/replate on Mon.
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:* Colony PCR of 3A assembly related colonies
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::* Colony #10 has potential. Will restreak on Mon.
== Drylab Work ==
== Drylab Work ==

Revision as of 21:54, 5 September 2008

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

Things we did today

Wetlab work

Construction of Broad-Host-Range Vector Intermediates

Margaret
  • Plasmid Prep: Everything that didn't work yesterday
  • 2 Plac + B0030, 1 Plac + B0034, aadA(BB)colony 5, pSB3k3, pSB1A3
  • Restriction Digest: same as Yesterday
  • Ligation
  • Transformation into DH5alpha (we need more competent cells)

Verification of transformants from yesterday

Grace
EtBr stained 2% agarose gel ran at 83V for 1.75 hours. Two microliters of PCR product were loaded into each well.
Construct Colonies
plac+rbs (no vector) 0
C0012 derived vector 12
C0012 derived vector, SAP treated 163
3A: plac+rbs+vector >300
  • BBpRL1338a related plates all had a lawn (no distinct colonies observed). Too low antibiotic conc.? Will attempt to restreak/replate on Mon.
  • Colony PCR of 3A assembly related colonies
  • Colony #10 has potential. Will restreak on Mon.

Drylab Work

Sequencing

Margaret
  • Analysis of Sequencing submitted (oriV1-4, 2 aadA(BB), 2 aadA (pRL))


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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