Team:Hawaii/Notebook/2008-07-17

From 2008.igem.org

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(Colony PCR)
(Quote of the Day)
 
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===PCR===
===PCR===
:<strong> Grace </strong>
:<strong> Grace </strong>
 +
[[Image:071608 PCR.jpg|thumb|right|250px|PCR products on an EtBr stained 4% agarose gel run at 119V for 27 min then 95V for 25 min. Ten microliters of PCR reactions were loaded into each well.]]
:* Colony PCR for GFP fusion brick and new pRL1383a to verify inserts
:* Colony PCR for GFP fusion brick and new pRL1383a to verify inserts
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::* BBa_J04430
::* BBa_J04430
::* BBa_J33207
::* BBa_J33207
-
::* BBa_R0010
+
::* BBa_R001
:* Ran all PCR products on a 4% gel to check  
:* Ran all PCR products on a 4% gel to check  
::* Gel ran funny -- loading dye ran 2x as fast as the DNA itself
::* Gel ran funny -- loading dye ran 2x as fast as the DNA itself
 +
:::* 119V too much to handle? Wavy bands
::* Poor ladder resolution on gel
::* Poor ladder resolution on gel
::* No bands seen for GFP fusion, pRL1383a, R0010
::* No bands seen for GFP fusion, pRL1383a, R0010
-
::* Incorrect bands for B0015 and J33207
+
::* Incorrect bands for B0015 (multiple bands, what are the two bands at the top?) and J33207 (size too small, stuff still in wells)
::* Will redo plasmid preps for R0010, B0015, J33207
::* Will redo plasmid preps for R0010, B0015, J33207
::* Will redo PCR reaction tomorrow for pRL1383a, GFP fusion, B0015, J33207, R0010
::* Will redo PCR reaction tomorrow for pRL1383a, GFP fusion, B0015, J33207, R0010
=== Plasmid Prep===
=== Plasmid Prep===
-
:<strong> Grace <strong>
+
:<strong> Grace </strong>
-
:* Inoculated 7 ml LB + amp<sub>100<sub> with single colony of B0015, R0010, J04430 for redo of plasmid prep tomorrow
+
:* Inoculated 7 ml LB + amp<sub>100</sub> with single colony of B0015, R0010, J04430 for redo of plasmid prep tomorrow
=== Restreak to Purify===
=== Restreak to Purify===
-
:<strong>Grace<strong>
+
:<strong>Grace</strong>
:* Restreaked GFP fusion and pRL1383a (MCS replaced) transformants to purify
:* Restreaked GFP fusion and pRL1383a (MCS replaced) transformants to purify
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:<strong> Margaret </strong>
:<strong> Margaret </strong>
-
== Drylab Work ==
 
-
 
-
===Name of Task===
 
-
:<strong> name of person/people who performed the task</strong>
 
-
 
-
:* Summary of task and what was done. Link to experiment for detailed notes if necessary.
 
-
:* e.g. read through papers, worked on proposal, etc.
 
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= Quote of the Day =
= Quote of the Day =
-
<blockquote>''History is the only laboratory we have in which to test the consequences of thought.'' - Étienne Gilson</blockquote>
+
At the team meeting today:
 +
 
 +
: "We have cookies, we can start."
 +
 
 +
 
 +
''Ten cookies later...''
 +
 
 +
 
 +
: "The more cookies I have, the better I think!"
 +
 
 +
 
 +
::::::- Dr. Callahan
{{Team:Hawaii/Footer}}
{{Team:Hawaii/Footer}}
__NOTOC__
__NOTOC__

Latest revision as of 04:53, 18 July 2008

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Things we did today

Wetlab work

PCR

Grace
PCR products on an EtBr stained 4% agarose gel run at 119V for 27 min then 95V for 25 min. Ten microliters of PCR reactions were loaded into each well.
  • Colony PCR for GFP fusion brick and new pRL1383a to verify inserts
  • Sequencing (25 mu;l reactions)
  • nir promoter
  • slr2016
  • pilA
  • BBa_B0015
  • BBa_B0024
  • BBa_B0030
  • BBa_C0012
  • BBa_E0040
  • BBa_J04430
  • BBa_J33207
  • BBa_R001
  • Ran all PCR products on a 4% gel to check
  • Gel ran funny -- loading dye ran 2x as fast as the DNA itself
  • 119V too much to handle? Wavy bands
  • Poor ladder resolution on gel
  • No bands seen for GFP fusion, pRL1383a, R0010
  • Incorrect bands for B0015 (multiple bands, what are the two bands at the top?) and J33207 (size too small, stuff still in wells)
  • Will redo plasmid preps for R0010, B0015, J33207
  • Will redo PCR reaction tomorrow for pRL1383a, GFP fusion, B0015, J33207, R0010

Plasmid Prep

Grace
  • Inoculated 7 ml LB + amp100 with single colony of B0015, R0010, J04430 for redo of plasmid prep tomorrow

Restreak to Purify

Grace
  • Restreaked GFP fusion and pRL1383a (MCS replaced) transformants to purify

pSB3K3 plasmid prep

Margaret


Discussion

Quote of the Day

At the team meeting today:

"We have cookies, we can start."


Ten cookies later...


"The more cookies I have, the better I think!"


- Dr. Callahan


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