Difference between revisions of "Team:Pittsburgh/Notebook"
| Line 25: | Line 25: | ||
<ul> | <ul> | ||
<li>Training begins</li> | <li>Training begins</li> | ||
| − | <li>Grow Top 10 competent cells.</li> | + | <li>Grow Top 10 <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#competent" target="_blank">competent cells</a>.</li> |
</ul> | </ul> | ||
<h2>Dry Lab</h2> | <h2>Dry Lab</h2> | ||
| Line 38: | Line 38: | ||
<h2>Wet Lab</h2> | <h2>Wet Lab</h2> | ||
<ul> | <ul> | ||
| − | <li>Test efficiency of competent cells</li> | + | <li>Test <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#competent" target="_blank">efficiency</a> of competent cells</li> |
</ul> | </ul> | ||
<h3>Cell Free Extract</h3> | <h3>Cell Free Extract</h3> | ||
| − | <ul><li>Test cell free extract reaction with T7-GFP plasmid</li></ul> | + | <ul><li>Test <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#cellfree" target="_blank">cell free extract reaction</a> with T7-GFP plasmid</li></ul> |
<h2>Dry Lab</h2> | <h2>Dry Lab</h2> | ||
<ul> | <ul> | ||
| Line 54: | Line 54: | ||
<h3>Reporter</h3> | <h3>Reporter</h3> | ||
<ul> | <ul> | ||
| − | <li>Transform T7 promoter, amilCP, and terminator</li> | + | <li><a href="https://2016.igem.org/Team:Pittsburgh/Protocols#transformations" target="_blank">Transform</a> T7 promoter, amilCP, and terminator</li> |
| − | <li>Begin assembly by ligating linearized T7 promoter and amilCP</li> | + | <li>Begin assembly by <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#ligation" target="_blank">ligating</a> linearized T7 promoter and amilCP</li> |
</ul> | </ul> | ||
<h2>Dry Lab</h2> | <h2>Dry Lab</h2> | ||
| Line 68: | Line 68: | ||
<h3>Reporter</h3> | <h3>Reporter</h3> | ||
<ul> | <ul> | ||
| − | <li>Transform T7-GFP plasmid, lacZ alpha fragment, and eGFP</li> | + | <li><a href="https://2016.igem.org/Team:Pittsburgh/Protocols#transformations" target="_blank">Transform</a> T7-GFP plasmid, lacZ alpha fragment, and eGFP</li> |
| − | <li>Send promising T7 promoter -- amilCP ligations to be sequenced</li> | + | <li>Send promising T7 promoter -- amilCP ligations to be <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#sequencing" target="_blank">sequenced</a></li> |
| − | <li>Perform double digest of T7 promoter and terminator from last week</li> | + | <li>Perform <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#digest" target="_blank">double digest</a> of T7 promoter and terminator from last week</li> |
| − | <li>Ligate double-digested T7 promoter to new reporters (lacZ and eGFP)</li> | + | <li><a href="https://2016.igem.org/Team:Pittsburgh/Protocols#ligation" target="_blank">Ligate</a> double-digested T7 promoter to new reporters (lacZ and eGFP)</li> |
</ul> | </ul> | ||
<h2>Dry Lab</h2> | <h2>Dry Lab</h2> | ||
| Line 84: | Line 84: | ||
<h3>Reporter</h3> | <h3>Reporter</h3> | ||
<ul> | <ul> | ||
| − | <li>Ligate T7 promoter -- amilCP construct to terminator</li> | + | <li><a href="https://2016.igem.org/Team:Pittsburgh/Protocols#ligation" target="_blank">Ligate</a> T7 promoter -- amilCP construct to terminator</li> |
| − | <li>Extract successful ligations of T7 promoter to eGFP</li> | + | <li><a href="https://2016.igem.org/Team:Pittsburgh/Protocols#extraction" target="_blank">Extract</a> successful ligations of T7 promoter to eGFP</li> |
| − | <li>Ligate T7 promoter -- eGFP construct to terminator</li> | + | <li><a href="https://2016.igem.org/Team:Pittsburgh/Protocols#ligation" target="_blank">Ligate</a> T7 promoter -- eGFP construct to terminator</li> |
</ul> | </ul> | ||
<h3>Cell Free Extract</h3> | <h3>Cell Free Extract</h3> | ||
Revision as of 15:05, 28 June 2016
Contact Us
Thank you to our sponsors!
Our weekly progress
Week 1: May 23 - May 27
Wet Lab
- Training begins
- Grow Top 10 competent cells.
Dry Lab
- Brainstorm genetic circuits for a thallium sensor
- Lab safety training
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Week 2: May 31 - June 3
Wet Lab
- Test efficiency of competent cells
Cell Free Extract
- Test cell free extract reaction with T7-GFP plasmid
Dry Lab
- Contact museums and summer programs for outreach opportunities
- Lab safety training
Back to Top
Week 3: June 6 - June 12
Wet Lab
Reporter
- Transform T7 promoter, amilCP, and terminator
- Begin assembly by ligating linearized T7 promoter and amilCP
Dry Lab
- Contact museums and summer programs for outreach opportunities
Back to Top
Week 4: June 13 - June 17
Wet Lab
Reporter
- Transform T7-GFP plasmid, lacZ alpha fragment, and eGFP
- Send promising T7 promoter -- amilCP ligations to be sequenced
- Perform double digest of T7 promoter and terminator from last week
- Ligate double-digested T7 promoter to new reporters (lacZ and eGFP)
Dry Lab
- TECBio, DiSCoBio, and Tissue Engineering Camp outreach opportunities set
Back to Top
Week 5: June 20 - June 26
Wet Lab
Reporter
- Ligate T7 promoter -- amilCP construct to terminator
- Extract successful ligations of T7 promoter to eGFP
- Ligate T7 promoter -- eGFP construct to terminator
Cell Free Extract
- Reaction volume can be reduced to 5 μL with 5 ng/μL of DNA
Toehold Switch
- Collins triggers activate the switches (both in plasmid form) to express LacZ
Dry Lab
- Reach out to teams to collaborate based on last year's projects
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