Team:UT Dallas/week9

From 2011.igem.org

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<h2>Week 9</h2>
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<h2>Week 8</h2>
<table>
<table>
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<tr><td valign='top'>9/6/11</td><td valign='top'>Incubation:<br>Germany Parts=[pHKToxR'MalE, pHKToxR'MalE/ZIP, pHKToxR'REI-T39K, pHKToxR'MalE/ZIP L267S, pHKToxR'HisJGCN] (FHK12)<br>
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<tr><td valign='top'>8/28/11</td><td valign='top'>Transformation (DH5-alpha): GFP-RBS-TetR-term<br>
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  CheY, PyeaR-GFP-RBS-TetR-term, scp-GFP-RBS-TetR-Term<br>
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Incubation of CheZ, CheZ*</td></tr>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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<tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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<tr><td valign='top'>9/7/11</td><td valign='top'> Glycerol Stock/Miniprep: [pHKToxR'MalE, pHKToxR'MalE/ZIP, pHKToxR'REI-T39K, pHKToxR'MalE/ZIP L267S, pHKToxR'HisJGCN]<br>
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<tr><td valign='top'>8/29/11</td><td valign='top'> Glycerol/Miniprep ctx-CheZ, ctx-CheZ*
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Incubation: pYeaR-GFP-RBS-TetR-Term, scp-GFP-RBS-Tetr-Term, AHL inducible Colicin, Pcst-RBS-LuxI-Term<br>
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Digestion: ToxR, FGFR
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Full Digestion/Gel Electrophoresis+Purification: FGFR<br>
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Incubation ToxR</td></tr>
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Ligation 10 Min: Vectors: FGFR, Inserts; ToxR<br>
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<tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Transformation: ToxR-FGFR
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<tr><td valign='top'>8/30/11</td><td valign='top'> Test Digestion: ctx-CheZ, ctx-Chez*, CheZ, CheZ*<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Gel Electrophoresis: ctx-CheZ, ctx-Chez*<br>
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<tr><td valign='top'>9/8/11</td><td valign='top'> Dilution of Pcst-RBS-:uxI-Term, AHL induible colicin<br>
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Incubation: ToxR</td></tr>
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Incubation: DATA COLLECTION=GFP quantification [(a) Psct-RBS-LuxI-Term vs. (b) AHL inducible colicin](DH5-alpha)  <br>
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<tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Glycerol Stock/Miniprep: Pyear-GFP-RBS-TetR-Term, scp-GFP-RBS-TetR-Term<br>
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<tr><td valign='top'>8/31/11</td><td valign='top'> Miniprep: ToxR<br>
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Test Digestion: Pyear-GFP-RBS-Term, scp-GFP-RBS-TetR-Term<br>
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Full Digestion: ToxR, pSB1C3<br>
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Colony PCR/ Gel Ectrophoresis: Toxr-FGFR
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Gel Electrophoresis/Purifcation: ToxR,GFP -> CheZ*, FGFR, RBS-TetR-Term<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Transformation: Ctx-CheZ*, ToxR-FGFR, GFP-RBS-TetR-Term</td></tr>
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<tr><td valign='top'>9/9/11</td><td valign='top'> Full Digestion: FGFR, RBS-GFP-TetR-Term, CheY, CheZ*, ctx-CheZ<br>
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<tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Test Digestion: Pyear-GFp-RBS-TetR-Term, scp-GFP-RBS-TetR-Term<br>
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<tr><td valign='top'>9/1/11</td><td valign='top'> Plating: k422007 CheY<br>
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Ligation 10 Min: Vectors: CheY, CheZ*, FGFR, RBS-GFP  Inserts: Ctx<br>
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Incubation: GFP-RBS-TetR-Term (Carb)<br>
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Transformation: FGFR, ToxR-FGFR, ctx-CheZ*-RBS-GFP, CheZ*, ctx-CheZ*<br>
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Incubation: E.Coli FHK12 - from Germany (Carb)<br>
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</td></tr><tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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Incubation: CheZ* (Chlora)</td></tr>
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<tr><td valign='top'>9/10/11</td><td valign='top'> Colony PCR: ToxR-FGFR, ctx-CheZ*, ctx-CheZ </td></tr>
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<tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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<tr><td valign='top'>9/2/11</td><td valign='top'> Plates Made: CarbR, ChloraR, CarbR/ChloraR<br>
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Broth Made: LB CarbR, LB Carb/ChloraR<br>
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Miniprep: GFP-RBS-TetR-Term, CheZ*<br>
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Glycerol Stock: GFP-RBS-TetR-Term<br>
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Made Competent Cells: FHK12 (Carb)<br>
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Test Digestion/Gel Electrophoresis: GFP-RBS-TetR-Term,<br>
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Full Digestion: CheZ*, FGFR<br>
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Ful Digestion: GFP-RBS-TetR-Term<br>
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Transformation: German Parts=[pHKToxR'MalE, pHKToxR'MalE/ZIP, pHKToxR'REI-T39K, pHKToxR<br>
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Plasmid stock: Diluted German parts (as above)<br>
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Ligation 10 Min: ctx, ToxR into CheZ*, FGFR respectively<br>
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Transformation: CheZ*, ctx-CheZ*, FGFR, ToxR-FGFR<br>
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Ligation Overnight: Vectors= GFP-RBS-TetR-Term, PyeaR, scp  <br>
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Inserts= ctx, GFP-RBS, TetR-Term</td></tr>
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<tr><td valign='top'>&nbsp;</td><td valign='top'>&nbsp;</td></tr>
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<tr><td valign='top'>9/3/11</td><td valign='top'> Transformation of the followings:<br>ctx-GFP-RBS-TetR-Term, PyeaR-GFP-RBS-TetR-Term, scp-GFP-RBS-TetR-Term</td>
</table>
</table>

Revision as of 00:54, 28 September 2011

Week 8

8/28/11Transformation (DH5-alpha): GFP-RBS-TetR-term
Incubation of CheZ, CheZ*
  
8/29/11 Glycerol/Miniprep ctx-CheZ, ctx-CheZ*

Digestion: ToxR, FGFR

Incubation ToxR
  
8/30/11 Test Digestion: ctx-CheZ, ctx-Chez*, CheZ, CheZ*

Gel Electrophoresis: ctx-CheZ, ctx-Chez*

Incubation: ToxR
  
8/31/11 Miniprep: ToxR

Full Digestion: ToxR, pSB1C3
Gel Electrophoresis/Purifcation: ToxR,GFP -> CheZ*, FGFR, RBS-TetR-Term

Transformation: Ctx-CheZ*, ToxR-FGFR, GFP-RBS-TetR-Term
  
9/1/11 Plating: k422007 CheY

Incubation: GFP-RBS-TetR-Term (Carb)
Incubation: E.Coli FHK12 - from Germany (Carb)

Incubation: CheZ* (Chlora)
  
9/2/11 Plates Made: CarbR, ChloraR, CarbR/ChloraR

Broth Made: LB CarbR, LB Carb/ChloraR
Miniprep: GFP-RBS-TetR-Term, CheZ*
Glycerol Stock: GFP-RBS-TetR-Term
Made Competent Cells: FHK12 (Carb)
Test Digestion/Gel Electrophoresis: GFP-RBS-TetR-Term,
Full Digestion: CheZ*, FGFR
Ful Digestion: GFP-RBS-TetR-Term
Transformation: German Parts=[pHKToxR'MalE, pHKToxR'MalE/ZIP, pHKToxR'REI-T39K, pHKToxR
Plasmid stock: Diluted German parts (as above)
Ligation 10 Min: ctx, ToxR into CheZ*, FGFR respectively
Transformation: CheZ*, ctx-CheZ*, FGFR, ToxR-FGFR
Ligation Overnight: Vectors= GFP-RBS-TetR-Term, PyeaR, scp

Inserts= ctx, GFP-RBS, TetR-Term
  
9/3/11 Transformation of the followings:
ctx-GFP-RBS-TetR-Term, PyeaR-GFP-RBS-TetR-Term, scp-GFP-RBS-TetR-Term
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