Difference between revisions of "Team:Pittsburgh/Notebook"
| Line 17: | Line 17: | ||
<li><a href="#Week4" class="table">Week 4: June 13 - June 17</a></li> | <li><a href="#Week4" class="table">Week 4: June 13 - June 17</a></li> | ||
<li><a href="#Week5" class="table">Week 5: June 20 - June 26</a></li> | <li><a href="#Week5" class="table">Week 5: June 20 - June 26</a></li> | ||
| + | <li><a href="#Week6" class="table">Week 6: June 27 - July 3</a></li> | ||
</ul> | </ul> | ||
</div> | </div> | ||
| Line 40: | Line 41: | ||
<li>Test <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#competent" target="_blank">efficiency</a> 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 <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#cellfree" target="_blank">cell free extract reaction</a> 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 88: | Line 89: | ||
<li><a href="https://2016.igem.org/Team:Pittsburgh/Protocols#ligation" target="_blank">Ligate</a> 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> |
<ul> | <ul> | ||
<li>Reaction volume can be reduced to 5 μL with 5 ng/μL of DNA</li> | <li>Reaction volume can be reduced to 5 μL with 5 ng/μL of DNA</li> | ||
| Line 103: | Line 104: | ||
<a href="#Top">Back to Top</a> | <a href="#Top">Back to Top</a> | ||
| + | |||
| + | <h1 class="nav"><a name="Week6" class="nav">Week 5: June 20 - June 26</a></h1> | ||
| + | <h2>Wet Lab</h2> | ||
| + | <h3>Reporter</h3> | ||
| + | <ul> | ||
| + | <li>Identify successful ligations to terminator for amilCP and eGFP consturcts using a <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#agarosegel" target="_blank">gel</a></li> | ||
| + | <li>Send correct plasmids for <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#sequencing" target="_blank">sequencing</a> for confirmation</li> | ||
| + | <li>Test plasmids in <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#cellfree" target="_blank">cell-free extract</a></li> | ||
| + | <li>amilCP does not produce color in cell-free reaction</li> | ||
| + | <li>eGFP produces fluorescence comparable to that from the Collins T7-GFP plasmid in cell-free reaction</li> | ||
| + | </ul> | ||
| + | <h3>Cell-Free Extract</h3> | ||
| + | <ul> | ||
| + | <li>Reaction volume can be reduced to 1 μL with 5 ng/μL of DNA</li> | ||
| + | </ul> | ||
| + | <h3>Toehold Switch</h3> | ||
| + | <ul> | ||
| + | <li>Collins plasmids express LacZ with 25 ng of switch</li> | ||
| + | <li>DNA oligos trigger Collins switches</li> | ||
| + | </ul> | ||
| + | <h2>Dry Lab</h2> | ||
| + | <ul> | ||
| + | <li>Work on outreach presentation for tissue engineering camp</li> | ||
| + | </ul> | ||
| + | <a href="https://static.igem.org/mediawiki/2016/5/59/T--Pittsburgh--NotebookWeek6.pdf" target="_blank">Week 6 Notebook</a><br> | ||
| + | <a href="#Top">Back to Top</a> | ||
| + | |||
</div> | </div> | ||
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Revision as of 15:59, 5 July 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
Back to Top
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
Back to Top
Week 5: June 20 - June 26
Wet Lab
Reporter
- Identify successful ligations to terminator for amilCP and eGFP consturcts using a gel
- Send correct plasmids for sequencing for confirmation
- Test plasmids in cell-free extract
- amilCP does not produce color in cell-free reaction
- eGFP produces fluorescence comparable to that from the Collins T7-GFP plasmid in cell-free reaction
Cell-Free Extract
- Reaction volume can be reduced to 1 μL with 5 ng/μL of DNA
Toehold Switch
- Collins plasmids express LacZ with 25 ng of switch
- DNA oligos trigger Collins switches
Dry Lab
- Work on outreach presentation for tissue engineering camp
Back to Top
