Glasgow/Wetlab

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[[Glasgow/Wetlab/Protocols|PROTOCOLS]]
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!align=center|[[Image:Uog.jpg]] ||    ||  [[Glasgow|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Main Page</font>]] || [[Glasgow/Modeling|<font face=georgia color=#3366CC size=4>Go To <br> Glasgow's <br> Modelling Page</font>]]
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[[Glasgow/Wetlab/References|REFERENCES]]
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{|cellspacing="6px" cellpadding="16" border="0" width="100%"
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|[https://2007.igem.org/Glasgow/Wetlab/Protocols <font face=georgia color=#3366CC size=5><b>Protocols</b></font>]
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|[https://2007.igem.org/Glasgow/Wetlab/References <font face=georgia color=#3366CC size=5><b>References</b></font>]
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|[https://2007.igem.org/Glasgow/Wetlab/Resources <font face=georgia color=#3366CC size=5><b>Resources</b></font>]
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|[https://2007.igem.org/Glasgow/Wetlab/Orders <font face=georgia color=#3366CC size=5><b>Sequences</b></font>]
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|[https://2007.igem.org/Glasgow/Wetlab/Biobricks <font face=georgia color=#3366CC size=5><b>Biobricks<br>Used</b></font>]
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|[https://2007.igem.org/Glasgow/Wetlab/Gels <font face=georgia color=#3366CC size=5><b>Gels</b></font>]
|}
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== Week 1 ==
 
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=== 03/07 ===
 
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Maija and Christine prepared LB broth and LB agar with Protocol 1.  Made 4 batches of plates, 2 with kanamycin and 2 with carbenicillin.
 
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=== 04/07 ===
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1 All wetlab researched BioBricks.
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2 Reporter constructs and mini-Tn5 stocks looked out.
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3 Streaked the following: P. putida PAW 340 pJAK14 Carb. plate
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Tn5 lux AB Carb. plate
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Mini-Tn5 lux AB Carb. plate
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P. Fluorescens NCIMB 9815 Carb. plate
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P. putida KT2440 LB
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JM109 pBluescript 5k+ Carb. plate
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''Click on a week number for more detailed lab book...''
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Mini-Tn5 Tc Carb. Plate
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pQF52 Carb. plate
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P. Fluorescens 9815 LB
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E. coli pJAK14 Km plate
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Il DntR in pOF52 Carb. plate
 
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pUCINR in Ω strain C Carb. plate
 
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pGLTUR Carb. plate
 
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Mini-Tn5 Kan Carb. plate
 
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Mini-Tn5 Sm/Sp Carb. plate
 
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Mini-Tn5 1cc2 Carb. plate
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E. coli Sa1 LB
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DmpR #24 Carb. plate
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!width="50%" | [https://2007.igem.org/Glasgow/Wetlab/Week1 <font face=georgia color=#3366CC size=7><b>Week 1</b></font>] <br>
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Mini-Tn5 lac 32 in E. coli 517 Carb. plate
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Mini-Tn5 Tc Carb. Plate
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Mini-Tn5 Cm Carb. plate
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!width="50%" |[https://2007.igem.org/Glasgow/Wetlab/Week2 <font face=georgia color=#3366CC size=7><b>Week 2</b></font>] <br>
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DmpR WT Carb. plate
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E.coli sm 10 pESD15 Tn5 GFP Carb. plate
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pUJ8 Carb. Plate
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*'''First selection of biobricks transformed.'''
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*'''Preparation of stocks.'''
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*'''Researched DmpR, XylR and DntR sensor systems.'''
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*'''Designed primers for DmpR, DntR, XylR and associated promoters.'''
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|- align="center"
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!width="50%" |[https://2007.igem.org/Glasgow/Wetlab/Week3 <font face=georgia color=#3366CC size=7><b>Week 3</b></font>] <br>
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!width="50%" |[https://2007.igem.org/Glasgow/Wetlab/Week4 <font face=georgia color=#3366CC size=7><b>Week 4</b></font>] <br>
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*'''BioBricks tested by digestion and PCR.'''
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*'''PCR with new primers associated with DntR and XylR.'''
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|
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* '''phzM and phzS cloned into TOPO vectors.'''
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*'''Primer design for 7 gene operon.'''
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*'''DntR cloned into TOPO vector.'''
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*'''Primer design for XylR and associated genes.'''
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*'''Death gene cloned into TOPO vector.'''
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*'''Primer design for death gene.'''
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|- align="center"
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!width="50%" |[https://2007.igem.org/Glasgow/Wetlab/Week5 <font face=georgia color=#3366CC size=7><b>Week 5</b></font>] <br>
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!width="50%" |[https://2007.igem.org/Glasgow/Wetlab/Week6 <font face=georgia color=#3366CC size=7><b>Week 6</b></font>] <br>
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|- valign=top
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*'''Suspected amplification of 7 gene operon.'''
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* '''phzM and phzS confirmed as present in TOPO vectors.'''
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*'''Confirmed presence of death gene plasmid in some vectors.'''
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*'''Miller assay performed with DntR system.'''
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*'''Yeast cells.'''
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*'''Biobricks made of phzM, phzS, and DntR.'''
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|- align="center"
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!width="50%" |[https://2007.igem.org/Glasgow/Wetlab/Week7 <font face=georgia color=#3366CC size=7><b>Week 7</b></font>] <br>
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!width="50%" |[https://2007.igem.org/Glasgow/Wetlab/Week8 <font face=georgia color=#3366CC size=7><b>Week 8</b></font>] <br>
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*'''Utilised new biobricks.'''
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*'''Site directed mutagenesis of phzM completed.'''
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*'''Site directed mutagenesis of phzS completed.'''
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|- align="center"
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!width="50%" |[https://2007.igem.org/Glasgow/Wetlab/Week9 <font face=georgia color=#3366CC size=7><b>Week 9</b></font>] <br>
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!width="50%" |[https://2007.igem.org/Glasgow/Wetlab/Week10 <font face=georgia color=#3366CC size=7><b>Week 10+</b></font>] <br>
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|- valign=top
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*'''phzD→phzG successfully cloned into TOPO vector.'''
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*'''Contains further work achieved.'''
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*'''XylR and Pr cloned into construction vectors.'''
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*'''XylR and Pr put next to responsive promoter and luciferase.'''
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|}
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----
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{|cellspacing="6px" cellpadding="16" border="0" width="100%"
 +
|- align=center
 +
|[https://2007.igem.org/Glasgow/Wetlab/Protocols <font face=georgia color=#3366CC size=5><b>Protocols</b></font>]
 +
|[https://2007.igem.org/Glasgow/Wetlab/References <font face=georgia color=#3366CC size=5><b>References</b></font>]
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|[https://2007.igem.org/Glasgow/Wetlab/Resources <font face=georgia color=#3366CC size=5><b>Resources</b></font>]
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|[https://2007.igem.org/Glasgow/Wetlab/Orders <font face=georgia color=#3366CC size=5><b>Orders</b></font>]
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|[https://2007.igem.org/Glasgow/Wetlab/Biobricks <font face=georgia color=#3366CC size=5><b>Biobricks<br>Used</b></font>]
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|[https://2007.igem.org/Glasgow/Wetlab/Gels <font face=georgia color=#3366CC size=5><b>Gels</b></font>]
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|}

Latest revision as of 11:54, 26 October 2007

Uog.jpg Back To
Glasgow's
Main Page 		Go To
Glasgow's
Modelling Page
Protocols References Resources Sequences Biobricks
Used 		Gels

Click on a week number for more detailed lab book...


Week 1
Week 2
  • First selection of biobricks transformed.
  • Preparation of stocks.
  • Researched DmpR, XylR and DntR sensor systems.
  • Designed primers for DmpR, DntR, XylR and associated promoters.
Week 3
Week 4
  • BioBricks tested by digestion and PCR.
  • PCR with new primers associated with DntR and XylR.
  • phzM and phzS cloned into TOPO vectors.
  • Primer design for 7 gene operon.
  • DntR cloned into TOPO vector.
  • Primer design for XylR and associated genes.
  • Death gene cloned into TOPO vector.
  • Primer design for death gene.
Week 5
Week 6
  • Suspected amplification of 7 gene operon.
  • phzM and phzS confirmed as present in TOPO vectors.
  • Confirmed presence of death gene plasmid in some vectors.
  • Miller assay performed with DntR system.
  • Yeast cells.
  • Biobricks made of phzM, phzS, and DntR.
Week 7
Week 8
  • Utilised new biobricks.
  • Site directed mutagenesis of phzM completed.
  • Site directed mutagenesis of phzS completed.


Week 9
Week 10+
  • phzD→phzG successfully cloned into TOPO vector.
  • Contains further work achieved.
  • XylR and Pr cloned into construction vectors.
  • XylR and Pr put next to responsive promoter and luciferase.
Protocols References Resources Orders Biobricks
Used 		Gels
Retrieved from "http://2007.igem.org/wiki/index.php/Glasgow/Wetlab"