Team:Northwestern/Notebook/Week10

From 2011.igem.org

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<img src="https://static.igem.org/mediawiki/2011/4/40/Notebook_banner4_2.gif" height = "70px" width="750px" style="opacity:1;filter:alpha(opacity=100)" alt="NU-igem banner"/ border="0">
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<div style="margin: -55px 0px 0px 80px;font:35px helvetica; color:#ffffff;">
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Notebook  <div style="margin: -40px 0px 0px 420px;font:35px helvetica; color:#444444;">    &nbsp; Week 10</div>
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<!--Week 10-->
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<!-- ---------------------Day 44 --------------------------------------- -->
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<html><div id="day44" style="margin: 0px 0px 0px 0px;"><img src="https://static.igem.org/mediawiki/2011/6/6b/Day_banner3.gif" height = "55px" width="450px" style="opacity:1;filter:alpha(opacity=100)" alt="day banner"/ border="0">
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<div style="margin: -40px 0px 18px 20px;font:24px helvetica;font-weight:550; color:#ffffff;"> Day 44 - Monday, August 15th 2011
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</div></div></html>
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*Miniprepped overnight cultures of the genomic promoter ligations.
 +
*PCRed the parts that were successfully sequenced because we didn’t have much DNA
 +
*Attempted to re-PCR the strep tag on. None of our transformations grew, even the ones where we used kinase, so we think the original PCR never worked.
 +
*Worked on hammering out the details of our human practices project
 +
*Developed a presentation of our math model
 +
*Began ligating our sequence confirmed PCR products onto the correct backbones.
 +
*Digested the RBS34+Parts minipreps from friday and ran them on a gel to confirm their validity.
 +
*Started a competent cell test
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<!-- ---------------------Day 45 --------------------------------------- -->
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<html><div id="day45" style="margin: 0px 0px 0px 0px;"><img src="https://static.igem.org/mediawiki/2011/6/6b/Day_banner3.gif" height = "55px" width="450px" style="opacity:1;filter:alpha(opacity=100)" alt="day banner"/ border="0">
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<div style="margin: -40px 0px 18px 20px;font:24px helvetica;font-weight:550; color:#ffffff;"> Day 45 - Tuesday, August 17th 2011
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</div></div></html>
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*Prepared for a PAGE gel tomorrow to asses LasR and RhlR activity
 +
*Planned for testing at the HTA lab
 +
*Transformed yesterday’s backbone correction ligations.
 +
*Digested the Genomic Promoter + RBS composites, and ligated them with reporter genes.
 +
*Worked on catching up the few remaining parts that were not successfully sequenced, such as CP+RBS+RFP and CP+RBS+LasR.
 +
<!-- ------------------------------------------------------------------- -->
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<!-- ---------------------Day 46 --------------------------------------- -->
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<html><div id="day46" style="margin: 0px 0px 0px 0px;"><img src="https://static.igem.org/mediawiki/2011/6/6b/Day_banner3.gif" height = "55px" width="470px" style="opacity:1;filter:alpha(opacity=100)" alt="day banner"/ border="0">
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<div style="margin: -40px 0px 18px 20px;font:24px helvetica;font-weight:550; color:#ffffff;"> Day 46 - Wednesday, August 17th 2011
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</div></div></html>
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*All the backbone transformations succeeded! There were both red (original backbone) and white (correct product) on all the colonies.
 +
*Presented our math model to the advisers
 +
*Met with Sara at the high throughput lab to plan our testing, decided to run a control sample tomorrow.
 +
*Started growing up CP->RBS->Receptor cells for a PAGE gel and then sonicated them.
 +
*Transformed all of the genomic promoter ligations
 +
*Started overnight cultures for testing
 +
*Started overnight cultures (70!) of the backbone correction ligations.
 +
*Made Amp/Kan plates (among others) so that we could try a double transformation with the seperate construct plasmids if we want to or if putting them on the same plasmid caused any problems.
 +
<!-- ------------------------------------------------------------------- -->
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<!-- ---------------------Day 47 --------------------------------------- -->
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<html><div id="day47" style="margin: 0px 0px 0px 0px;"><img src="https://static.igem.org/mediawiki/2011/6/6b/Day_banner3.gif" height = "55px" width="450px" style="opacity:1;filter:alpha(opacity=100)" alt="day banner"/ border="0">
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<div style="margin: -40px 0px 18px 20px;font:24px helvetica;font-weight:550; color:#ffffff;"> Day 47 - Thursday, August 18th 2011
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</div></div></html>
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*Completed 70 minipreps of our backbone correction ligations!
 +
*Ran a PAGE gel to asses LasR and RhlR activity
 +
*Made new SOB and plates
 +
*Our cells were not growing properly in the m9 media, so we made it again from a different recipe. This has forced us to delay our testing.
 +
*Analyzed a recently published paper (yesterday!) that provides a lot of relevant information about our project.
 +
*Ligated our strep tag parts
 +
*Stored the genomic promoter transformations in the cold room. They looked good, but we are postponing overnights to catch up on everything else.
 +
<!-- ------------------------------------------------------------------- -->
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<!-- ---------------------Day 48 --------------------------------------- -->
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<html><div id="day48" style="margin: 0px 0px 0px 0px;"><img src="https://static.igem.org/mediawiki/2011/6/6b/Day_banner3.gif" height = "55px" width="450px" style="opacity:1;filter:alpha(opacity=100)" alt="day banner"/ border="0">
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<div style="margin: -40px 0px 18px 20px;font:24px helvetica;font-weight:550; color:#ffffff;"> Day 48 - Friday, August 19th 2011
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</div></div></html>
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*We digested all 70 minipreps from yesterday with the X and P enzymes. In the afternoon, we ran all 70 digests on gels to confirm that our constructs have been correctly inserted into their final backbones.
 +
*We got the results of our PAGE gel
 +
*We tested samples to calibrate the differences between our spectrophotometer and the High Throughput Lab plate reader.
 +
*We finished remaking M9 media.
 +
*Transformed Strep Tag Ligations

Latest revision as of 03:42, 23 September 2011

RETURN TO IGEM 2010



day banner
Day 44 - Monday, August 15th 2011

  • Miniprepped overnight cultures of the genomic promoter ligations.
  • PCRed the parts that were successfully sequenced because we didn’t have much DNA
  • Attempted to re-PCR the strep tag on. None of our transformations grew, even the ones where we used kinase, so we think the original PCR never worked.
  • Worked on hammering out the details of our human practices project
  • Developed a presentation of our math model
  • Began ligating our sequence confirmed PCR products onto the correct backbones.
  • Digested the RBS34+Parts minipreps from friday and ran them on a gel to confirm their validity.
  • Started a competent cell test


day banner
Day 45 - Tuesday, August 17th 2011

  • Prepared for a PAGE gel tomorrow to asses LasR and RhlR activity
  • Planned for testing at the HTA lab
  • Transformed yesterday’s backbone correction ligations.
  • Digested the Genomic Promoter + RBS composites, and ligated them with reporter genes.
  • Worked on catching up the few remaining parts that were not successfully sequenced, such as CP+RBS+RFP and CP+RBS+LasR.


day banner
Day 46 - Wednesday, August 17th 2011

  • All the backbone transformations succeeded! There were both red (original backbone) and white (correct product) on all the colonies.
  • Presented our math model to the advisers
  • Met with Sara at the high throughput lab to plan our testing, decided to run a control sample tomorrow.
  • Started growing up CP->RBS->Receptor cells for a PAGE gel and then sonicated them.
  • Transformed all of the genomic promoter ligations
  • Started overnight cultures for testing
  • Started overnight cultures (70!) of the backbone correction ligations.
  • Made Amp/Kan plates (among others) so that we could try a double transformation with the seperate construct plasmids if we want to or if putting them on the same plasmid caused any problems.


day banner
Day 47 - Thursday, August 18th 2011

  • Completed 70 minipreps of our backbone correction ligations!
  • Ran a PAGE gel to asses LasR and RhlR activity
  • Made new SOB and plates
  • Our cells were not growing properly in the m9 media, so we made it again from a different recipe. This has forced us to delay our testing.
  • Analyzed a recently published paper (yesterday!) that provides a lot of relevant information about our project.
  • Ligated our strep tag parts
  • Stored the genomic promoter transformations in the cold room. They looked good, but we are postponing overnights to catch up on everything else.


day banner
Day 48 - Friday, August 19th 2011

  • We digested all 70 minipreps from yesterday with the X and P enzymes. In the afternoon, we ran all 70 digests on gels to confirm that our constructs have been correctly inserted into their final backbones.
  • We got the results of our PAGE gel
  • We tested samples to calibrate the differences between our spectrophotometer and the High Throughput Lab plate reader.
  • We finished remaking M9 media.
  • Transformed Strep Tag Ligations
Retrieved from "http://2011.igem.org/Team:Northwestern/Notebook/Week10"