Team:UT Dallas/week12

From 2011.igem.org

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<h2>Week 11</h2>
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<h2>Week 12</h2>
<table>
<table>
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<tr><td valign='top'>9/11/11</td><td valign='top'>Incubation: ToxR-FGFR, ctx-CheZ*, ctx-CheZ, ToxR, FGFR, CheY, CheZ,
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<tr><td valign='top'>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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9/18/11</td><td valign='top'> Miniprep/Glycerol Stock: pYeaR-ToxR-FGFR, ctx-CheZ*, CheY<br>
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<tr><td valign='top'>9/12/11</td><td valign='top'>Glycerol Stock: ToxR-FGFR, ctx-CheZ*, ctx-CheZ, ToxR, FGFR, CheY, CheZ,<br>
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Test Digestion: pYeaR-ToxR-FGFR, ctx-CheZ*, CheY <br>
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Test Digestion/Gel Electrophoresis: FGFR-ToxR, ctx-CheZ-RBS-GFP<br>
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Colony PCR: pYeaR-ToxR-FGFR<br>
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Full Digestion/Gel Electrophoresis: FGFR-ToxR, ctx-CheZ-RBS-GFP<br>
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Test Digestion: CheY (X/S)
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Transformation: ToxR'MalE + ctx-CheZ-RBS-GFP, ToxR'MalE/ZIP + ctx-CheZ-RBS-GFP, ToxR'MalE/Zip L267S + ctx-CheZ-RBS-GFP<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr><tr><td valign='top'>
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Plating: ToxR'MalE/ZIP + ctx-CheZ-RBS-GFP<br>
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9/19/11</td><td valign='top'> Full Digestion/Electrophoresis: CheY, CheZ*, pSB1C3<br>
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Ligation: scp, Pyear, TetR, ToxR-FGFR, ctx<br>
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PCR/Gel Electrophoresis: CheY<br>
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Incubation: CheZ* (Chlora)
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Ligation: ctx + CheZ*, ctx + CheY, pYeaR-ToxR-FGFR + pSB1C3, ctx-CheZ-RBS-GFP + pSB1C3<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Incubation: scp-ToxR-FGFR, ToxR'MalE/ZIP, ToxR'MalE
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<tr><td valign='top'>9/13/11</td><td valign='top'>Transform of PyeaR-ToxR-FGFR, TetR-ToxR-FGFR, scp-ToxR-FGFR<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr><tr><td valign='top'>
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Full Digestion: ToxR-FGFR (at E/X as vector)<br>
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9/20/11</td><td valign='top'> PCR/Gel Purify: ctx<br>
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Ligation: Vector-> ToxR-FGFR  Insert-> ctx<br>
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Maltose Induction Experiment: DATA COLLECTION<br>
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Incubation: CheY
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Miniprep: scp-ToxR-FGFR<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Full Digestion/Gel Purification: CheZ*, CheY<br>
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<tr><td valign='top'>9/14/11</td><td valign='top'>Transformation: ctx-ToxR-FGFR<br>
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Ligation: ctx + CheZ*, ctx + CheY, ctx + CheZ-RBS-GFP<br>
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Test Digestion: ToxR'MalE, ToxR'MalE/ZIP, ToxR'MalE/Zip L267S<br>
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Transformation: ToxR'MalE + ctx-GFP, ToxR'MalE/ZIP + ctx-GFP, ToxR'MalE/ZIP L267S + ctx-GFP<br>
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Incubation: scp-ToxR-FGFR, TetR-ToxR-FGFR, PyeaR-ToxR-FGFR (Carb)<br>
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Icubation: pYeaR-ToxR-FGFR (Chlora)<br>
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Miniprep: CheY<br>
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Miniprep/Glycerol Stock: pYeaR-ToxR-FGFR
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PCR: CheY<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr><tr><td valign='top'>
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Incubation: ToxR'MalE, ToxR'MalE/ZIP, CheZ*
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9/21/11</td><td valign='top'> PCR: ctx (retry)<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Test Digestion/Purification: pYeaR-ToxR-FGFR<br>
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<tr><td valign='top'>9/15/11</td><td valign='top'>PCR: CheY<br>
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Full Digestion/PurificationL pYeaR-ToxR-FGFR<br>
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Full Digestion: CheY (X/S), vector pSB1C3<br>
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Dephosphorylation: pSB1C3<br>
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Miniprep/Glycerol Stock: pYeaR-ToxR-FGFR, scp-ToxR-FGFR, ToxR'MalE, ToxR'MalE/ZIP<br>
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Incubation: ctx-gfp, BL21 E. coli<br>
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Test Digestion: pYeaR-ToxR-FGFR, scp-ToxR-FGFR, ToxR'MalE, ToxR'MalE/ZIP<br>
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Transformation: Germany Parts (FHK12)<br>
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Dephosphorylation: Vector pSB1C3<br>
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Gel Purification: ctx<br>
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Ligation: CheY + pSB1c3<br>
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Ligation: ctx + CheY<br>
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Full Digestion/Gel Elctrophoresis/Purification: pYeaR-ToxR-FGFR, scp-ToxR-FGFR, ToxR'MalE, ToxR'MalE/ZIP, CheZ, CheZ*, ctx-CheZ-RBS-GFP, ctx<br>
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Incubation: ctx-CheZ*
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Ligation: ctx + CheZ*<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr><tr><td valign='top'>
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Transformation: pSB1C3 Vector, pYeaR-ToxR-FGFR, ctx-CheZ-RBS-GFP, CheZ, ctx-GFP, ctx, Chey
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9/22/11</td><td valign='top'> Miniprep/Glycerol Stock: ctx-GFP, ctx-CheZ, ctx-CheY<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Full Digestion: pSB1C3<br>
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<tr><td valign='top'>9/16/11</td><td valign='top'>Digestion: ctx-CheZ<br>
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Make BL21 competent<br>
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Incubation: ToxR'MalE/ZIP, ToxR'MalE, <br>
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Dephosphorylation: pSB1C3<br>
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Ligation: ctx + CheY (retry)<br>
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Ligation: ctx + CheY, ctx + FGFR-ToxR<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Transformation: (BL21) pcst-RBS-LuxI, AHL inducible colicin, ctx-CheY, ctx-ToxR-FGFR<br>
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<tr><td valign='top'>9/17/11</td><td valign='top'>Incubation: pYeaR-ToxR-FGFR, ctx-CheZ*, CheY</td></tr>
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Ligation: scp-ToxR-GFP + pSB1C3, ctx-CheZ-RBS-GFP + pSB1C3, ctx-Chez + pSB1C3<br>
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Incubation: psct-RBS-LuxI-term
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr><tr><td valign='top'>
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9/23/11</td><td valign='top'> Miniprep: psct-RBS-LuxI-term<br>
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Dissolving/Diluting Primers: Germany Parts<br>
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1:20 Dilution of scp-ToxR-FGFR + ctx-GFP, pYeaR-ToxR-FGFR + ctx-GFP <br>
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Glycerol Stock: scp-ToxR-FGFR + ctx-GFP, pYeaR-ToxR-FGFR + ctx-GFP <br>
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Transformation: German Parts<br>
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Incubation: scp-ToxR-FGFR, pYeaR-ToxR-FGFR
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr><tr><td valign='top'>
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9/24/11</td><td valign='top'> Dilution 1:10 for Optical Density readings: scp-ToxR-FGFR, pYeaR-ToxR-FGFR<br>
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Growing from glycerol stock (backup): scp-ToxR-FGFR, pYeaR-ToxR-FGFR<br>
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PCR: German Parts<br>
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Digestion: pSB1C3 (X/S)  <br>
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Heparin/FGF Induction Experiment: DATA COLLECTION (Fluorescence)<br>
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Dephosphorylation: Germany Parts, pSB1C3<br>
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Ligation: <br> &nbsp;&nbsp;&nbsp;&nbsp;ctx-CheZ + pSB1C3, scp-ToxR-FGFR + pSB1C3, ctx-CheZ-RBS-GFP + pSB1C3, pSB1C3-CheY + ctx, pSB1C3-CheZ* + ctx, pSb1C3-ToxR-FGFR + ctx<br>
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Transformation: <br>&nbsp;&nbsp;&nbsp;&nbsp;ctx-CheZ + pSB1C3, scp-ToxR-FGFR + pSB1C3, ctx-CheZ-RBS-GFP + pSB1C3, pSB1C3-CheY + ctx, pSB1C3-CheZ* + ctx, pSb1C3-ToxR-FGFR + ctx
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Transformation: <br>&nbsp;&nbsp;&nbsp;&nbsp;ToxR'MalE, ToxR'MalE/ZIP, ToxR'MalE/ZIP L267S</td></tr>
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<tr><td valign='top'>
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9/25/11</td><td valign='top'> Dilution 1:20 for experimentation: ToxR'MalE, ToxR'MalE/ZIP, ToxR'MalE/ZIP L267S <br>
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Running Maltose Induction Experiment: DATA COLLECTION (fluorescence)   <br>
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Incubation: scp-ToxR-FGFR, pYeaR-ToxR-FGFR, <br>
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Incubation of fluorescent parts: scp-ToxR-FGFR, pYeaR-ToxR-FGFR,
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr><tr><td valign='top'>
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9/26/11</td><td valign='top'> Dilution 1:5 for experimentation: pYear-ToxR-FGFR, scp-ToxR-FGFR   <br>
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Heparin/FGF Induction Experiment (re-run): DATA COLLECTION (fluorescence)    <br>
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Incubation (for iGem Registry): FGFR, CheZ*, ToxR-FGFR</td></tr>  
</table>
</table>

Revision as of 00:53, 28 September 2011

Week 12

9/18/11 Miniprep/Glycerol Stock: pYeaR-ToxR-FGFR, ctx-CheZ*, CheY

Test Digestion: pYeaR-ToxR-FGFR, ctx-CheZ*, CheY
Colony PCR: pYeaR-ToxR-FGFR
Test Digestion: CheY (X/S)

  
9/19/11 Full Digestion/Electrophoresis: CheY, CheZ*, pSB1C3

PCR/Gel Electrophoresis: CheY
Ligation: ctx + CheZ*, ctx + CheY, pYeaR-ToxR-FGFR + pSB1C3, ctx-CheZ-RBS-GFP + pSB1C3
Incubation: scp-ToxR-FGFR, ToxR'MalE/ZIP, ToxR'MalE

  
9/20/11 PCR/Gel Purify: ctx

Maltose Induction Experiment: DATA COLLECTION
Miniprep: scp-ToxR-FGFR
Full Digestion/Gel Purification: CheZ*, CheY
Ligation: ctx + CheZ*, ctx + CheY, ctx + CheZ-RBS-GFP
Transformation: ToxR'MalE + ctx-GFP, ToxR'MalE/ZIP + ctx-GFP, ToxR'MalE/ZIP L267S + ctx-GFP
Icubation: pYeaR-ToxR-FGFR (Chlora)
Miniprep/Glycerol Stock: pYeaR-ToxR-FGFR

  
9/21/11 PCR: ctx (retry)

Test Digestion/Purification: pYeaR-ToxR-FGFR
Full Digestion/PurificationL pYeaR-ToxR-FGFR
Dephosphorylation: pSB1C3
Incubation: ctx-gfp, BL21 E. coli
Transformation: Germany Parts (FHK12)
Gel Purification: ctx
Ligation: ctx + CheY
Incubation: ctx-CheZ*

  
9/22/11 Miniprep/Glycerol Stock: ctx-GFP, ctx-CheZ, ctx-CheY

Full Digestion: pSB1C3
Make BL21 competent
Dephosphorylation: pSB1C3
Ligation: ctx + CheY, ctx + FGFR-ToxR
Transformation: (BL21) pcst-RBS-LuxI, AHL inducible colicin, ctx-CheY, ctx-ToxR-FGFR
Ligation: scp-ToxR-GFP + pSB1C3, ctx-CheZ-RBS-GFP + pSB1C3, ctx-Chez + pSB1C3
Incubation: psct-RBS-LuxI-term

  
9/23/11 Miniprep: psct-RBS-LuxI-term

Dissolving/Diluting Primers: Germany Parts
1:20 Dilution of scp-ToxR-FGFR + ctx-GFP, pYeaR-ToxR-FGFR + ctx-GFP
Glycerol Stock: scp-ToxR-FGFR + ctx-GFP, pYeaR-ToxR-FGFR + ctx-GFP
Transformation: German Parts
Incubation: scp-ToxR-FGFR, pYeaR-ToxR-FGFR

  
9/24/11 Dilution 1:10 for Optical Density readings: scp-ToxR-FGFR, pYeaR-ToxR-FGFR

Growing from glycerol stock (backup): scp-ToxR-FGFR, pYeaR-ToxR-FGFR
PCR: German Parts
Digestion: pSB1C3 (X/S)
Heparin/FGF Induction Experiment: DATA COLLECTION (Fluorescence)
Dephosphorylation: Germany Parts, pSB1C3
Ligation:
    ctx-CheZ + pSB1C3, scp-ToxR-FGFR + pSB1C3, ctx-CheZ-RBS-GFP + pSB1C3, pSB1C3-CheY + ctx, pSB1C3-CheZ* + ctx, pSb1C3-ToxR-FGFR + ctx
Transformation:
    ctx-CheZ + pSB1C3, scp-ToxR-FGFR + pSB1C3, ctx-CheZ-RBS-GFP + pSB1C3, pSB1C3-CheY + ctx, pSB1C3-CheZ* + ctx, pSb1C3-ToxR-FGFR + ctx

Transformation:
    ToxR'MalE, ToxR'MalE/ZIP, ToxR'MalE/ZIP L267S
9/25/11 Dilution 1:20 for experimentation: ToxR'MalE, ToxR'MalE/ZIP, ToxR'MalE/ZIP L267S

Running Maltose Induction Experiment: DATA COLLECTION (fluorescence)
Incubation: scp-ToxR-FGFR, pYeaR-ToxR-FGFR,
Incubation of fluorescent parts: scp-ToxR-FGFR, pYeaR-ToxR-FGFR,

  
9/26/11 Dilution 1:5 for experimentation: pYear-ToxR-FGFR, scp-ToxR-FGFR

Heparin/FGF Induction Experiment (re-run): DATA COLLECTION (fluorescence)

Incubation (for iGem Registry): FGFR, CheZ*, ToxR-FGFR
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