Team:Northwestern/Notebook/Week10
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+ | <div id="header" style="margin: 14px 0px 0px 0px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2011/4/40/Notebook_banner4_2.gif" height = "auto" width="750px" style="opacity:1;filter:alpha(opacity=100)" alt="NU-igem banner"/ border="0"> | ||
+ | <div style="margin: -55px 0px 0px 80px;font:35px helvetica; color:#ffffff;"> | ||
+ | Notebook <div style="margin: -40px 0px 0px 420px;font:35px helvetica; color:#444444;"> Week 10</div> | ||
+ | </div></div> | ||
+ | </html> | ||
+ | <br> | ||
+ | |||
+ | |||
+ | <!--Week 10--> | ||
+ | |||
+ | <!-- ---------------------Day 44 --------------------------------------- --> | ||
+ | <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"> | ||
+ | <div style="margin: -40px 0px 18px 20px;font:24px helvetica;font-weight:550; color:#ffffff;"> Day 44 - Monday, August 15th 2011 | ||
+ | </div></div></html> | ||
+ | *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 45 --------------------------------------- --> | ||
+ | <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"> | ||
+ | <div style="margin: -40px 0px 18px 20px;font:24px helvetica;font-weight:550; color:#ffffff;"> Day 45 - Tuesday, August 17th 2011 | ||
+ | </div></div></html> | ||
+ | *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 46 --------------------------------------- --> | ||
+ | <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"> | ||
+ | <div style="margin: -40px 0px 18px 20px;font:24px helvetica;font-weight:550; color:#ffffff;"> Day 46 - Wednesday, August 17th 2011 | ||
+ | </div></div></html> | ||
+ | *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 47 --------------------------------------- --> | ||
+ | <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"> | ||
+ | <div style="margin: -40px 0px 18px 20px;font:24px helvetica;font-weight:550; color:#ffffff;"> Day 47 - Thursday, August 18th 2011 | ||
+ | </div></div></html> | ||
+ | *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 48 --------------------------------------- --> | ||
+ | <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"> | ||
+ | <div style="margin: -40px 0px 18px 20px;font:24px helvetica;font-weight:550; color:#ffffff;"> Day 48 - Friday, August 19th 2011 | ||
+ | </div></div></html> | ||
+ | *Remember to transform strep tag ligations! |
Revision as of 14:48, 19 August 2011
PROJECT
RESULTS
CONSIDERATIONS
ABOUT US
NOTEBOOK
ATTRIBUTIONS
Notebook
Week 10
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 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 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 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 48 - Friday, August 19th 2011
- Remember to transform strep tag ligations!