Team:Hawaii/Notebook/2008-10-23

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(Difference between revisions)
(→Large scale plasmid prep)
(→Large scale plasmid prep)
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:*From 9/28 sp<sub>2.5</sub> plate: #1
:*From 9/28 sp<sub>2.5</sub> plate: #1
::* Gel showed no DNA in lanes migrated. Huh? Will try one more time to plasmid prep tomorrow.
::* Gel showed no DNA in lanes migrated. Huh? Will try one more time to plasmid prep tomorrow.
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 +
===Inoculated for plasmid prep===
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:<strong>Grace</strong>
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:* In 10 ml TB+amp<sub>100</sub>:
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::* nir
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::* nir+rbs
 +
::* slr1
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::* pilA
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::* GFPf
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::* slr1+GFPf
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:* In 50 ml TB+sp<sub>100</sub>:
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::* BB1 (sp<sub>100</sub> plate)
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:* Incubated at 37C for 18 hours with shaking at 250 rpm
===Verification of transformants===
===Verification of transformants===

Revision as of 22:35, 24 October 2008

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Things we did today

Wetlab work

Antibiotic test

Grace
  • No change from yesterday
  • Colony PCR of 6 colonies from sp40, 50, 75 and 8 colonies from sp100 plates
  • Restreaked sp2.5 blue colony, sp100 #1 & 2 on LB+amp100, LB, LB+sp100 + X-gal

Large scale plasmid prep

Grace
  • From sp100 plate: #1, 2
  • From 9/28 sp2.5 plate: #1
  • Gel showed no DNA in lanes migrated. Huh? Will try one more time to plasmid prep tomorrow.

Inoculated for plasmid prep

Grace
  • In 10 ml TB+amp100:
  • nir
  • nir+rbs
  • slr1
  • pilA
  • GFPf
  • slr1+GFPf
  • In 50 ml TB+sp100:
  • BB1 (sp100 plate)
  • Incubated at 37C for 18 hours with shaking at 250 rpm

Verification of transformants

EtBr stained 2% agarose gel ran at 95V for 1 hour. Two microliters of PCR product were loaded into each lane.
Grace
Colony count of transformants
Plasmid Colonies
J04430 0
(-) on amp100 0
BB-1 4
(-) on sp100 0
  • Colony PCR of 4 BB-1 transformants
  • None were correct

RE digest

Grace
  • EcoRI/PstI: nir+rbs
  • XbaI/SpeI: slr1+GFPf

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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