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Team:Northwestern/Notebook/Week8
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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"> | ||
| + | <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 8</div> | ||
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<!--Week 8--> | <!--Week 8--> | ||
| - | < | + | <!-- ---------------------Day 34 --------------------------------------- --> |
| - | Day 34 - | + | <html><div id="day34" 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 34 - Monday, August 1st 2011 | |
| - | ---------------------- | + | </div></div></html> |
*Miniprepped constitutive promoters, RBSs, and a 3 part ligation | *Miniprepped constitutive promoters, RBSs, and a 3 part ligation | ||
*Digested 2A (w/ PCR clean-up) and 3A for ligations of RBSs with parts | *Digested 2A (w/ PCR clean-up) and 3A for ligations of RBSs with parts | ||
| Line 14: | Line 24: | ||
*Found strep tag sequence? Fingers crossed. Worked on primers for adding tag to receptors. | *Found strep tag sequence? Fingers crossed. Worked on primers for adding tag to receptors. | ||
*Prepared samples for sequencing (lasP+RBS30+RFP, rhlP+RBS34+GFP) | *Prepared samples for sequencing (lasP+RBS30+RFP, rhlP+RBS34+GFP) | ||
| + | <!-- ------------------------------------------------------------------- --> | ||
| - | < | + | <!-- ---------------------Day 35 --------------------------------------- --> |
| - | Day 35 - | + | <html><div id="day35" 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 35 - Tuesday, August 2nd 2011 | |
| - | ---------------------- | + | </div></div></html> |
*Made glycerol stock of constitutive promoter + RBS34 + rhlR | *Made glycerol stock of constitutive promoter + RBS34 + rhlR | ||
*Digested, ligated, and transformed constitutive promoters and RBSs/parts | *Digested, ligated, and transformed constitutive promoters and RBSs/parts | ||
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*Used up chlor plates for transformations so we made more plates (chlor and tet) | *Used up chlor plates for transformations so we made more plates (chlor and tet) | ||
*Plated ligations from glycerol stocks | *Plated ligations from glycerol stocks | ||
| + | <!-- ------------------------------------------------------------------- --> | ||
| - | < | + | <!-- ---------------------Day 36 --------------------------------------- --> |
| - | Day 36 - | + | <html><div id="day36" 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 36 - Wednesday, August 3rd 2011 | |
| - | ---------------------- | + | </div></div></html> |
*Miniprepped yesterday’s ONs | *Miniprepped yesterday’s ONs | ||
*Made glycerol stock of CP2 | *Made glycerol stock of CP2 | ||
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*Digested RBSs+parts and ligated with CP oligo | *Digested RBSs+parts and ligated with CP oligo | ||
*Most of yesterday’s transforms did not work - ONed transforms that did work | *Most of yesterday’s transforms did not work - ONed transforms that did work | ||
| + | <!-- ------------------------------------------------------------------- --> | ||
| - | < | + | <!-- ---------------------Day 37 --------------------------------------- --> |
| - | Day 37 - | + | <html><div id="day37" 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 37 - Thursday, August 4th 2011 | |
| - | ---------------------- | + | </div></div></html> |
*Miniprepped, digested, and ran gels of RBS34+parts and RBS30+parts ligations | *Miniprepped, digested, and ran gels of RBS34+parts and RBS30+parts ligations | ||
*Miniprepped chlor backbone | *Miniprepped chlor backbone | ||
| Line 50: | Line 63: | ||
*Plated RBS30+rhlR from glycerol stock | *Plated RBS30+rhlR from glycerol stock | ||
*Plated TOP10 cells seedstock in preparation for making more competent cells | *Plated TOP10 cells seedstock in preparation for making more competent cells | ||
| + | <!-- ------------------------------------------------------------------- --> | ||
| - | < | + | <!-- ---------------------Day 38 --------------------------------------- --> |
| - | Day 38 - | + | <html><div id="day38" 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 38 - Friday, August 5th 2011 | |
| - | ---------------------- | + | </div></div></html> |
*Made new sterile MgCl2 | *Made new sterile MgCl2 | ||
*Made glycerol stocks of what worked on yesterday’s gel | *Made glycerol stocks of what worked on yesterday’s gel | ||
*Worked on our super awesome iGEM t-shirt design | *Worked on our super awesome iGEM t-shirt design | ||
Latest revision as of 03:43, 23 September 2011
PROJECT
RESULTS
CONSIDERATIONS
ABOUT US
NOTEBOOK
ATTRIBUTIONS
Notebook
Week 8
Day 34 - Monday, August 1st 2011
- Miniprepped constitutive promoters, RBSs, and a 3 part ligation
- Digested 2A (w/ PCR clean-up) and 3A for ligations of RBSs with parts
- Ran gel of digested 3 part ligation
- Transformed cells with ligated digestions
- Found strep tag sequence? Fingers crossed. Worked on primers for adding tag to receptors.
- Prepared samples for sequencing (lasP+RBS30+RFP, rhlP+RBS34+GFP)
Day 35 - Tuesday, August 2nd 2011
- Made glycerol stock of constitutive promoter + RBS34 + rhlR
- Digested, ligated, and transformed constitutive promoters and RBSs/parts
- Finished primers for adding tag to receptors.
- Prepared samples for sequencing.
- PCRed a bunch of parts and 2L ligations; did ONs of backbones...we’re running out of DNA! =O
- Received promoter oligos in mail!
- Used up chlor plates for transformations so we made more plates (chlor and tet)
- Plated ligations from glycerol stocks
Day 36 - Wednesday, August 3rd 2011
- Miniprepped yesterday’s ONs
- Made glycerol stock of CP2
- Annealed CP oligos received in mail
- Digested RBSs+parts and ligated with CP oligo
- Most of yesterday’s transforms did not work - ONed transforms that did work
Day 37 - Thursday, August 4th 2011
- Miniprepped, digested, and ran gels of RBS34+parts and RBS30+parts ligations
- Miniprepped chlor backbone
- Transformed oligo ligations
- Plated RBS30+rhlR from glycerol stock
- Plated TOP10 cells seedstock in preparation for making more competent cells
Day 38 - Friday, August 5th 2011
- Made new sterile MgCl2
- Made glycerol stocks of what worked on yesterday’s gel
- Worked on our super awesome iGEM t-shirt design
