Team:Hawaii/Notebook/2008-10-23

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(→Verification of transformants)
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:* No change from yesterday
:* No change from yesterday
:* Colony PCR of 6 colonies from sp<sub>40, 50, 75</sub> and 8 colonies from sp<sub>100</sub> plates
:* Colony PCR of 6 colonies from sp<sub>40, 50, 75</sub> and 8 colonies from sp<sub>100</sub> plates
 +
::* sp<sub>75,100</sub> colonies correct
 +
:* Restreaked sp<sub>2.5</sub> blue colony, sp<sub>100</sub> #1 & 2 on LB+amp<sub>100</sub>, LB, LB+sp<sub>100</sub> + X-gal
 +
 +
===Large scale plasmid prep===
 +
:<strong>Grace</strong>
 +
 +
:*From sp<sub>100</sub> plate: #1, 2
 +
:*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.
 +
 +
===Inoculated for plasmid prep===
 +
:<strong>Grace</strong>
 +
:* In 10 ml TB+amp<sub>100</sub>:
 +
::* nir
 +
::* nir+rbs
 +
::* slr1
 +
::* pilA
 +
::* GFPf
 +
::* slr1+GFPf
 +
:* In 50 ml TB+sp<sub>100</sub>:
 +
::* BB1 (sp<sub>100</sub> plate)
 +
:* Incubated at 37C for 18 hours with shaking at 250 rpm
===Verification of transformants===
===Verification of transformants===
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|align=center|0
|align=center|0
|-
|-
-
|align=center|BB-1
+
|align=center|BB-1 (plasmid from 9/28 plate)
|align=center|4
|align=center|4
|-
|-
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|}
|}
:* Colony PCR of 4 BB-1 transformants
:* Colony PCR of 4 BB-1 transformants
 +
::* None were correct
 +
 +
===RE digest===
 +
:<strong>Grace</strong>
 +
 +
:* EcoRI/PstI: nir+rbs
 +
:* XbaI/SpeI: slr1+GFPf
 +
 +
===Plasmid Prep===
 +
:<strong>Krystle</strong>
 +
:*Plasmid prep of
 +
:**lac+rbs+slr+gfpf #1,3,4
 +
:**nir+rbs+slr+gfpf #2,3 (from last week)
= Discussion =
= Discussion =

Latest revision as of 01:07, 30 October 2008

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

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
  • sp75,100 colonies correct
  • 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 (plasmid from 9/28 plate) 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

Plasmid Prep

Krystle
  • Plasmid prep of
    • lac+rbs+slr+gfpf #1,3,4
    • nir+rbs+slr+gfpf #2,3 (from last week)

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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