Team:Northwestern/Notebook/Week9

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 9</div>
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<!--Week 9-->
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<!-- ---------------------Day 39 --------------------------------------- -->
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<html><div id="day39" 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 39 - Monday, August 8th 2011
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</div></div></html>
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*Miniprepped overnight cultures from the attempted Oligo based ligations last week.
 +
*Digested the minipreps and ran them on a gel to look for the correct sized bands.
 +
*Prepared samples for sequencing
 +
*Made new Tet Plates, Chlor Plates and SOB Broth
 +
*PCRed our strep tag primer onto our CP->LasR and CP-> RhlR constructs
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<!-- ------------------------------------------------------------------- -->
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<!-- ---------------------Day 40 --------------------------------------- -->
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<html><div id="day40" 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 40 - Tuesday, August 9th 2011
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</div></div></html>
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*Made a new batch of competent cells. We tried to get them in the -80 freezer extremely quickly to improve the efficiency.
 +
*We digested our strep tag PCR products with DpnI and then added ligase so the blunt ends would come back together. We ligated one batch for one hour and another overnight.
 +
*We transformed the overnight ligations
 +
*We transformed to test the competency of our new competent cells
 +
*Sent in samples for seqencing
 +
*Designed our T-shirts!
 +
*Rafay gave a presentation on the math model
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<!-- ------------------------------------------------------------------- -->
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<!-- ---------------------Day 41 --------------------------------------- -->
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<html><div id="day41" 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 41 - Wednesday, August 10th 2011
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</div></div></html>
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*Our competent cell test had only 21 colonies, indicating a very low efficiency =(
 +
*Our transformations of the strep tag ligations also failed, although we can still try transforming the overnight ligations.
 +
*Hopefully we’ll get sequencing results soon....
 +
*Miniprepped the overnights from our glycerol stocks and of our terminators
 +
*Transformed the strep tag overnight ligations from yesterday
 +
<!-- ------------------------------------------------------------------- -->
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<!-- ---------------------Day 42 --------------------------------------- -->
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<html><div id="day42" 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 42 - Thursday, August 11th 2011
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</div></div></html>
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*The overnight ligations of the strep tag constructs produced four colonies! These were  overnighted for miniprep tomorrow.
 +
*Ran another digest of the CP -> RBS -> LasR/RhlR -> Strep tag constructs, this time after phosphorylating the ends of the DNA. This should help improve ligation efficiency.
 +
*Ligated those digestions overnight
 +
*Ran our strep tag PCR products on a gel to try and confirm that the initial amplification was successful.
 +
*Prepared overnights of RBS+Part ligations from glycerol stocks and transformation plates. We will ligate our genomic promoters to these tomorrow.
 +
*Started 3 test ligations with Genomic Promoter -> RBS -> Part from already existing stocks.
 +
*Made a lot of progressing designing our human practices project
 +
*The team and sponsorship pages on the wiki are nearing completion.
 +
<!-- ------------------------------------------------------------------- -->
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<!-- ---------------------Day 43 --------------------------------------- -->
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<html><div id="day43" 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 43 - Friday, August 12th 2011
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</div></div></html>
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*Transformed the strep tag ligations
 +
*Miniprepped overnights of RBS34+Part Ligations
 +
*We got our sequencing results back! Most of the samples look very good and closely match what we expected.
 +
*Made some more progress on the website.

Latest revision as of 03:43, 23 September 2011

RETURN TO IGEM 2010



day banner
Day 39 - Monday, August 8th 2011

  • Miniprepped overnight cultures from the attempted Oligo based ligations last week.
  • Digested the minipreps and ran them on a gel to look for the correct sized bands.
  • Prepared samples for sequencing
  • Made new Tet Plates, Chlor Plates and SOB Broth
  • PCRed our strep tag primer onto our CP->LasR and CP-> RhlR constructs


day banner
Day 40 - Tuesday, August 9th 2011

  • Made a new batch of competent cells. We tried to get them in the -80 freezer extremely quickly to improve the efficiency.
  • We digested our strep tag PCR products with DpnI and then added ligase so the blunt ends would come back together. We ligated one batch for one hour and another overnight.
  • We transformed the overnight ligations
  • We transformed to test the competency of our new competent cells
  • Sent in samples for seqencing
  • Designed our T-shirts!
  • Rafay gave a presentation on the math model


day banner
Day 41 - Wednesday, August 10th 2011

  • Our competent cell test had only 21 colonies, indicating a very low efficiency =(
  • Our transformations of the strep tag ligations also failed, although we can still try transforming the overnight ligations.
  • Hopefully we’ll get sequencing results soon....
  • Miniprepped the overnights from our glycerol stocks and of our terminators
  • Transformed the strep tag overnight ligations from yesterday


day banner
Day 42 - Thursday, August 11th 2011

  • The overnight ligations of the strep tag constructs produced four colonies! These were overnighted for miniprep tomorrow.
  • Ran another digest of the CP -> RBS -> LasR/RhlR -> Strep tag constructs, this time after phosphorylating the ends of the DNA. This should help improve ligation efficiency.
  • Ligated those digestions overnight
  • Ran our strep tag PCR products on a gel to try and confirm that the initial amplification was successful.
  • Prepared overnights of RBS+Part ligations from glycerol stocks and transformation plates. We will ligate our genomic promoters to these tomorrow.
  • Started 3 test ligations with Genomic Promoter -> RBS -> Part from already existing stocks.
  • Made a lot of progressing designing our human practices project
  • The team and sponsorship pages on the wiki are nearing completion.


day banner
Day 43 - Friday, August 12th 2011

  • Transformed the strep tag ligations
  • Miniprepped overnights of RBS34+Part Ligations
  • We got our sequencing results back! Most of the samples look very good and closely match what we expected.
  • Made some more progress on the website.
Retrieved from "http://2011.igem.org/Team:Northwestern/Notebook/Week9"