Difference between revisions of "Team:Pittsburgh/Notebook"
| Line 9: | Line 9: | ||
</div> | </div> | ||
| − | <div | + | <div class="table column full_size"> |
| − | <h2><a class="nav">Contents</a></h2> | + | <h2><a name="Top" class="nav">Contents</a></h2> |
<ul class="table"> | <ul class="table"> | ||
<li><a href="#Week1" class="table">Week 1: May 23 - May 27</a></li> | <li><a href="#Week1" class="table">Week 1: May 23 - May 27</a></li> | ||
| Line 23: | Line 23: | ||
<div class="notebook column full_size"> | <div class="notebook column full_size"> | ||
| − | + | <h1 class="nav"><a name="Week1" class="nav">Week 1: May 23 - May 27</a></h1> | |
| − | <h1 class="nav"><a class="nav">Week 1: May 23 - May 27</a></h1> | + | |
<h2>Wet Lab</h2> | <h2>Wet Lab</h2> | ||
<ul> | <ul> | ||
Revision as of 17:20, 10 July 2016
Contact Us
Thank you to our sponsors!
Our weekly progress. For a list of our protocols, visit the Protocols page
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 6: June 27 - July 3
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
- Linearized plasmids containing only the promoter and insert (no terminator) do not express protein
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
Week 7: July 5 - July 8
Wet Lab
Reporter
- Sequenced amilCP construct does not contain amilCP
- Unsuccessfully linearize and amplify eGFP construct using PCR
Cell-Free Extract
- 384-well plate requires at least 10 μL of reaction
DNAzyme
- Anneal PO strand with catalytic strand, both with and without erbium
- Test success of annealing reaction in cell-free extract and with acrylamide gels
Dry Lab
- Practice outreach presentation for tissue engineering camp
- Develop DNAzymes for other heavy metals
Back to Top
