iGEM 2016 — Team Pittsburgh

[![](https://static.igem.org/mediawiki/2016/8/8b/HQ_page_logo.jpg)](https://2016.igem.org)

# Team:Pittsburgh/Notebook

[![](https://static.igem.org/mediawiki/2016/2/2e/T--Pittsburgh--templateBanner.jpg)](/Team%3APittsburgh)

* [Home](https://2016.igem.org/Team%3APittsburgh)
* People

  [Team](https://2016.igem.org/Team%3APittsburgh/Team)
  [Collaborations](https://2016.igem.org/Team%3APittsburgh/Collaborations)
   [InterLab](https://2016.igem.org/Team%3APittsburgh/InterLab)
   [Attributions](https://2016.igem.org/Team%3APittsburgh/Attributions)
* [Medals](https://2016.igem.org/Team%3APittsburgh/Medals)
* Project

  [Overview](https://2016.igem.org/Team%3APittsburgh/Project_Overview)
   [Experiments](https://2016.igem.org/Team%3APittsburgh/Experiments)
   [Results](https://2016.igem.org/Team%3APittsburgh/Results)
  [Proof of Concept](https://2016.igem.org/Team%3APittsburgh/Proof)
   [Notebook](https://2016.igem.org/Team%3APittsburgh/Notebook)
   [Protocols](https://2016.igem.org/Team%3APittsburgh/Protocols)
* Parts

  [Parts](https://2016.igem.org/Team%3APittsburgh/Parts)
   [Basic Parts](https://2016.igem.org/Team%3APittsburgh/Basic_Part)
   [Description](https://2016.igem.org/Team%3APittsburgh/Description)
* [Safety](https://2016.igem.org/Team%3APittsburgh/Safety)
* Human Practices

  [Human Practices](https://2016.igem.org/Team%3APittsburgh/Human_Practices)
  [Silver](https://2016.igem.org/Team%3APittsburgh/HP/Silver)
  [Gold](https://2016.igem.org/Team%3APittsburgh/HP/Gold)
  [Engagement](https://2016.igem.org/Team%3APittsburgh/Engagement)
  [Model](https://2016.igem.org/Team%3APittsburgh/Model)

#

[Home](/Team%3APittsburgh)

People

[Team](/Team%3APittsburgh/Team)

[Collaborations](/Team%3APittsburgh/Collaborations)

[InterLab](/Team%3APittsburgh/InterLab)

[Attributions](/Team%3APittsburgh/Attributions)

[Medals](/Team%3APittsburgh/Medals)

Project

[Overview](/Team%3APittsburgh/Project_Overview)

[Experiments](/Team%3APittsburgh/Experiments)

[Results](/Team%3APittsburgh/Results)

[Proof of Concept](/Team%3APittsburgh/Proof)

[Notebook](/Team%3APittsburgh/Notebook)

[Protocols](/Team%3APittsburgh/Protocols)

Parts

[Parts](/Team%3APittsburgh/Parts)

[Basic Part](/Team%3APittsburgh/Basic_Part)

[Description](/Team%3APittsburgh/Description)

[Safety](/Team%3APittsburgh/Safety)

Human Practices

[Human Practices](/Team%3APittsburgh/Human_Practices)

[Silver](/Team%3APittsburgh/HP/Silver)

[Gold](/Team%3APittsburgh/HP/Gold)

[Engagement](https://2016.igem.org/Team%3APittsburgh/Engagement)

[Model](/Team%3APittsburgh/Model)

Contact Us

pitt.igem.2016@gmail.com

Thank you to our sponsors!

[![Office of the Provost](https://static.igem.org/mediawiki/2016/4/46/T--Pittsburgh--SponsorsProvost.png)](http://www.provost.pitt.edu/)
[![Department of Bioengineering](https://static.igem.org/mediawiki/2016/4/4a/T--Pittsburgh--SponsorsSwanson.jpg)](http://www.engineering.pitt.edu/bioengineering/)
[![Department of Biological Sciences](https://static.igem.org/mediawiki/2016/5/53/T--Pittsburgh--SponsorsBiology.jpg)](http://www.biology.pitt.edu/)
[![Department of Chemistry](https://static.igem.org/mediawiki/2016/a/a7/T--Pittsburgh--SponsorsChemistry.jpg)](http://www.chem.pitt.edu/)
 [![Swanson School of Engineering](https://static.igem.org/mediawiki/2016/9/9e/T--Pittsburgh--SponsorsSwansonEngineering.png)](http://www.engineering.pitt.edu/)

[![IDT](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--SponsorsIDT.jpg)](https://www.idtdna.com/site)
[![NEB](https://static.igem.org/mediawiki/2016/4/44/T--Pittsburgh--SponsorsNEB.jpg)](https://www.neb.com/)
[![Experiment](https://static.igem.org/mediawiki/2016/6/67/T--Pittsburgh--templateExperiment.png)](https://experiment.com/)

Our weekly activities: experiments, data analysis, and planning. For a list of our protocols, visit the  [Protocols](https://2016.igem.org/Team%3APittsburgh/Protocols) page.

## Contents

* [Week 1: May 23 - May 27](#Week1)
* [Week 2: May 31 - June 3](#Week2)
* [Week 3: June 6 - June 12](#Week3)
* [Week 4: June 13 - June 17](#Week4)
* [Week 5: June 20 - June 26](#Week5)
* [Week 6: June 27 - July 3](#Week6)
* [Week 7: July 5 - July 8](#Week7)
* [Week 8: July 11 - July 17](#Week8)
* [Week 9: July 18 - July 22](#Week9)
* [Week 10: July 25 - July 31](#Week10)
* [Week 11: August 1 - August 5](#Week11)
* [Week 12: August 8 - August 13](#Week12)
* [Week 13: August 15 - August 20](#Week13)
* [Week 14: August 23 - August 26](#Week14)
* [Week 15: August 29 - September 2](#Week15)
* [Week 16: September 6 - September 9](#Week16)
* [Week 17: September 12 - September 17](#Week17)
* [Week 18: September 19 - September 24](#Week18)
* [Week 19: September 28 - September 30](#Week19)
* [Week 20: October 3 - October 8](#Week20)
* [Week 21: October 10 - October 14](#Week21)
* [Week 22: October 17 - October 19](#Week22)
* [Week 23: October 24 - October 31](#Week23)

# Week 1: May 23 - May 27

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

* Training begins
* Grow Top 10 [competent cells](https://2016.igem.org/Team%3APittsburgh/Protocols#competent).

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Brainstorm genetic circuits for a thallium sensor
* [Lab safety training](2016.igem.org/Team%3APittsburgh/Safety)

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 1 Notebook](https://static.igem.org/mediawiki/2016/b/bc/TeamPittsburghNotebookWeek1.pdf)
[Back to Top](#Top)

# Week 2: May 31 - June 3

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

* Test [efficiency](https://2016.igem.org/Team%3APittsburgh/Protocols#competent) of competent cells

### Cell-Free Reactions

* Test [cell-free extract reaction](https://2016.igem.org/Team%3APittsburgh/Protocols#cellfree) with T7-GFP plasmid

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Contact museums and summer programs for outreach opportunities
* [Lab safety training](2016.igem.org/Team%3APittsburgh/Safety)

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 2 Notebook](https://static.igem.org/mediawiki/2016/a/a8/TeamPittsburghNotebookWeek2.pdf)
[Back to Top](#Top)

# Week 3: June 6 - June 12

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### Reporter

* [Transform](https://2016.igem.org/Team%3APittsburgh/Protocols#transformations) T7 promoter, amilCP, and terminator
* Begin assembly by [ligating](https://2016.igem.org/Team%3APittsburgh/Protocols#ligation) linearized T7 promoter and amilCP

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Contact museums and summer programs for outreach opportunities

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 3 Notebook](https://static.igem.org/mediawiki/2016/b/b3/TeamPittsburghNotebookWeek3.pdf)
[Back to Top](#Top)

# Week 4: June 13 - June 17

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### Reporter

* [Transform](https://2016.igem.org/Team%3APittsburgh/Protocols#transformations) T7-GFP plasmid, lacZ alpha fragment, and eGFP
* Send promising T7 promoter -- amilCP ligations to be [sequenced](https://2016.igem.org/Team%3APittsburgh/Protocols#sequencing)
* Perform [double digest](https://2016.igem.org/Team%3APittsburgh/Protocols#digest) of T7 promoter and terminator from last week
* [Ligate](https://2016.igem.org/Team%3APittsburgh/Protocols#ligation) double-digested T7 promoter to new reporters (lacZ and eGFP)

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* TECBio, DiSCoBio, and Camp BioE outreach opportunities set

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 4 Notebook](https://static.igem.org/mediawiki/2016/7/75/TeamPittsburghNotebookWeek4.pdf)
[Back to Top](#Top)

# Week 5: June 20 - June 26

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### Cell-Free Reactions

* 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

### Reporter

* [Ligate](https://2016.igem.org/Team%3APittsburgh/Protocols#ligation) T7 promoter -- amilCP construct to terminator
* [Extract](https://2016.igem.org/Team%3APittsburgh/Protocols#extraction) successful ligations of T7 promoter to eGFP
* [Ligate](https://2016.igem.org/Team%3APittsburgh/Protocols#ligation) T7 promoter -- eGFP construct to terminator

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Reach out to teams to collaborate based on last year's projects

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 5 Notebook](https://static.igem.org/mediawiki/2016/7/7c/TeamPittsburghNotebookWeek5.pdf)
[Back to Top](#Top)

# Week 6: June 27 - July 3

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### Cell-Free Reactions

* 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

### Reporter

* Identify successful ligations to terminator for amilCP and eGFP constructs using a [gel](https://2016.igem.org/Team%3APittsburgh/Protocols#agarosegel)
* Send correct plasmids for [sequencing](https://2016.igem.org/Team%3APittsburgh/Protocols#sequencing) for confirmation
* Test plasmids in [cell-free extract](https://2016.igem.org/Team%3APittsburgh/Protocols#cellfree)
* 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

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Work on outreach presentation for  [Camp BioE](https://2016.igem.org/Team%3APittsburgh/Human_Practices)

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 6 Notebook](https://static.igem.org/mediawiki/2016/5/59/T--Pittsburgh--NotebookWeek6.pdf)
[Back to Top](#Top)

# Week 7: July 5 - July 8

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### Cell-Free Reactions

* 384-well plate requires at least 10 μL of reaction

### DNAzyme

* [Anneal](https://2016.igem.org/Team%3APittsburgh/Protocols#annealing) PO strand with catalytic strand, both with and without erbium
* Test success of annealing reaction in cell-free extract and with acrylamide gels

### Reporter

* Sequenced amilCP construct does not contain amilCP
* Unsuccessfully linearize and amplify eGFP construct using [PCR](https://2016.igem.org/Team%3APittsburgh/Protocols#pcr)

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Practice outreach presentation for [Camp BioE](https://2016.igem.org/Team%3APittsburgh/Human_Practices)
* Develop DNAzymes for other heavy metals

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 7 Notebook](https://static.igem.org/mediawiki/2016/d/d7/T--Pittsburgh--NotebookWeek7.pdf)
[Back to Top](#Top)

# Week 8: July 11 - July 17

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### Cell-Free Reactions

* Linear eGFP construct does not produce a stronger signal than its plasmid form

### DNAzyme

* DNAzyme duplex does not trigger toehold switch
* Erbium cleaves the P substrate strand

### Reporter

* Restart amilCP cloning process

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* First presentation at  [Camp BioE](https://2016.igem.org/Team%3APittsburgh/Human_Practices)
* Prepare for  [UMD Mid-Atlantic Meet-Up](https://2016.igem.org/Team%3APittsburgh/Collaborations#UMD)
* Contact PLSG and NEB for sponsorship

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 8 Notebook](https://static.igem.org/mediawiki/2016/c/cb/T--Pittsburgh--NotebookWeek8.pdf)
[Back to Top](#Top)

# Week 9: July 18 - July 22

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### Cell-Free Reactions

* Reactions can be diluted by one-half and still produce visible results in two hours

### DNAzyme

* dPAGE assay of P substrate cleavage suggests that the DNAzyme works, but results are not definitive
* Six-hour time course of cleavage does not yield much additional information
* Reaction temperature (room temperature versus 37°C) does not produce observable effect on cleavage rates

### Reporter

* Continue amilCP cloning process

### Amplifier

* Clone RBS-T3 RNA polymerase to add into other constructs

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Presentation at  [Camp BioEhttps://2016.igem.org/Team:Pittsburgh/Collaborations#UMD](https://2016.igem.org/Team%3APittsburgh/Human_Practices)
* [UMD Mid-Atlantic Meet-Up](https://2016.igem.org/Team%3APittsburgh/Collaborations#UMD)
* Continue fundraising
* Discuss systems to model

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 9 Notebook](https://static.igem.org/mediawiki/2016/7/75/T--Pittsburgh--NotebookWeek9.pdf)
[Back to Top](#Top)

# Week 10: July 25 - July 31

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### Cell-Free Reactions

* Reactions diluted by one-half produce significantly less protein than undiluted reactions

### DNAzyme

* Annealing reactions produce hybrid complexes but leave unsequestered substrate strand
* DNAzyme does not cleave P substrate strand

### Reporter

* Continue cloning amilCP construct
* Determine sequence of possible *lacZ* plasmid
* Unsuccessfully grow *lacZ* from iGEM bacterial stab

### Amplifier

* Continue cloning T3 constructs

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Presentation at  [Camp BioE](https://2016.igem.org/Team%3APittsburgh/Human_Practices)
* Start modeling toehold kinetics and the economical effects of lead

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 10 Notebook](https://static.igem.org/mediawiki/2016/6/64/T--Pittsburgh--NotebookWeek10.pdf)
[Back to Top](#Top)

# Week 11: August 1 - August 5

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

* Obtain fluorescence readings for [UGA Archaeal InterLab](/Team%3APittsburgh/Collaborations#UGA)

### Cell-Free Reactions

* Presence of erbium and Buffer B as part of cleavage reaction does not seem to affect reaction progress

### DNAzyme

* Higher catalytic-to-substrate strand ratios help increase sequestration of substrate strand, especially for G switch
* Cell-free reactions suggest that cleavage of the P substrate strand does occur in the presence of erbium
* dPAGE assay does not suggest cleavage

### Reporter

* amilCP construct contains CFP, not amilCP
* Contact Collins lab for PT3-GFP construct

### Amplifier

* Obtain T3 RNA polymerase gene via amplification
* Ligate PT3 and PT3-RBS into plasmid backbone

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Presentation at  [Camp BioE](https://2016.igem.org/Team%3APittsburgh/Human_Practices)
* Meeting with Dr. Daniel Bain from the University of Pittsburgh Department of Geology and Environmental Science
* Model population effects of lead without economic layer

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 11 Notebook](https://static.igem.org/mediawiki/2016/9/9f/T--Pittsburgh--NotebookWeek11.pdf)
[Back to Top](#Top)

# Week 12: August 8 - August 13

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### DNAzyme

* Begin work with lead DNAzyme
* dPAGE assay does not suggest cleavage occurs under the current conditions

### Reporter

* Unsuccessful mutagensis of *lacZ* to remove EcoRI site for BioBricking

### Amplifier

* Successfully ligate PT3-RBS into plasmid backbone
* Clone PT3-RBS-T3 and PT7-RBS-T3

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* [Table at Carnegie Science Center's H2Oh! Exhibit](https://2016.igem.org/Team%3APittsburgh/Human_Practices)
* Work on Experiment.com video for crowdfunding

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 12 Notebook](https://static.igem.org/mediawiki/2016/5/57/T--Pittsburgh--NotebookWeek12.pdf)
[Back to Top](#Top)

# Week 13: August 15 - August 20

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

* Complete data collection for  [InterLab study](/Team%3APittsburgh/InterLab)
* Prepare William & Mary plasmids for cell-free expression

### DNAzyme

* Start working with hairpin lead DNAzyme
* Cell-free reactions suggest that a hairpin DNAzyme cleaves more efficiently than a DNAzyme in a duplex
* Adding the DNAzyme, substrate, and metal to a cell-free reaction produces greater switch activation
   than adding a completed cleavage reaction
* Cell-free reactions suggest that cleavage efficiency is similar at 37°C and room temperature

### Reporter

* Unsuccessful mutagenesis of *lacZ* with DMSO

### Amplifier

* Unsuccessfully ligate terminator onto PT3-T3 and PT7-T3 constructs

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Work on Experiment.com video for crowdfunding

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 13 Notebook](https://static.igem.org/mediawiki/2016/8/8a/T--Pittsburgh--NotebookWeek13.pdf)
[Back to Top](#Top)

# Week 14: August 23 - August 26

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

* Complete data collection for William & Mary (read our report [here](https://static.igem.org/mediawiki/2016/4/46/T--Pittsburgh--CollaborationsWM_report.pdf))

### Amplifier

* ligate terminator onto PT3-T3 and PT7-T3 constructs

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Meet with Dr. Troesken from Pitt's Department of Economics to discuss lead population model

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 14 Notebook](https://static.igem.org/mediawiki/2016/2/23/T--Pittsburgh--NotebookWeek14.pdf)
[Back to Top](#Top)

# Week 15: August 29 - September 2

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### Cell-Free Reactions

* No reactions, including the positive control, turn purple to indicate the presence of LacZ

### Reporter

* Unsuccessful mutagenesis of *lacZ*

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Move lab up to campus

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 15 Notebook](https://static.igem.org/mediawiki/2016/f/f7/T--Pittsburgh--NotebookWeek15.pdf)
[Back to Top](#Top)

# Week 16: September 6 - September 9

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

* Obtain codon-optimized OFP, GFP, BFP, YFP, and RFP from 2015 Carnegie Mellon team

### Cell-Free Reactions

* Plasmids from last week's reactions did not actually contain the toehold switch

### DNAzyme

* Replicate results from Week 13 suggesting that hairpin lead DNAzyme sequesters and cleaves as expected

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Experiment.com campaign approved

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 16 Notebook](https://static.igem.org/mediawiki/2016/7/79/T--Pittsburgh--NotebookWeek16.pdf)
[Back to Top](#Top)

# Week 17: September 12 - September 17

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### DNAzyme

* Presence of lead produces signal higher than background hairpin activation at 3.74 nM DNAzyme
* Presence of lead may produce signal beyond background activation at 50 nM and 100 nM DNAzyme

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Experiment.com campaign is live

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 17 Notebook](https://static.igem.org/mediawiki/2016/4/4d/T--Pittsburgh--NotebookWeek17.pdf)
[Back to Top](#Top)

# Week 18: September 19 - September 24

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### DNAzyme

* Presence of lead produces signal higher than background hairpin activation at 3.74 nM DNAzyme
* Presence of lead does not produce signal beyond background activation at 50 nM and 100 nM DNAzyme

### Reporter

* Unsuccessful mutagenesis of *lacZ*

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Modeling

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 18 Notebook](https://static.igem.org/mediawiki/2016/5/5a/T--Pittsburgh--NotebookWeek18.pdf)
[Back to Top](#Top)

# Week 19: September 28 - September 30

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### DNAzyme

* PAGE analysis suggests cleavage of hairpin but is not clear
* Presence of lead does not produce signal beyond background activation at 50 nM and 100 nM DNAzyme

### Reporter

* Unsuccessful mutagenesis of *lacZ*

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Modeling

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 19 Notebook](https://static.igem.org/mediawiki/2016/4/48/T--Pittsburgh--NotebookWeek19.pdf)
[Back to Top](#Top)

# Week 20: October 3 - October 8

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

### DNAzyme

* PAGE analysis suggests cleavage of hairpin but is not clear

### Reporter

* Unsuccessful mutagenesis of *lacZ*

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 19 Notebook](https://static.igem.org/mediawiki/2016/6/62/T--Pittsburgh--NotebookWeek20.pdf)
[Back to Top](#Top)

# Week 21: October 10 - October 14

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

* Characterize codon-optimized fluorescent proteins from 2015 Carnegie Mellon team

### Cell-Free Reactions

* Expected dosage response to lead not observed
* Expected lack of activation with D switch not observed
* Reaction colors do not look the same as in previous weeks

### Reporter

* Unsuccessful mutagenesis of *lacZ*

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Modeling
* Finalize Wiki

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 21 Notebook](https://static.igem.org/mediawiki/2016/c/cc/T--Pittsburgh--NotebookWeek21.pdf)
[Back to Top](#Top)

# Week 22: October 17 - October 19

![Wet Lab](https://static.igem.org/mediawiki/2016/b/b8/T--Pittsburgh--NotebookWetLab.jpg)

* Finish cloning Parts
* Submit Parts

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Finalize Wiki

![notebook](https://static.igem.org/mediawiki/2016/9/96/T--Pittsburgh--NotebookNotebook.png)[Week 22 Notebook](https://static.igem.org/mediawiki/2016/f/f2/T--Pittsburgh--NotebookWeek22.pdf)
[Back to Top](#Top)

# Week 23: October 24 - October 31

![Dry Lab](https://static.igem.org/mediawiki/2016/e/e0/T--Pittsburgh--NotebookDryLab.jpg)

* Prepare poster and presentation
* Giant Jamboree!

[Back to Top](#Top)

[![](https://static.igem.org/mediawiki/2016/b/bf/HQ_menu_logo.jpg)](https://2016.igem.org)