Glasgow/Wetlab/Week1
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| - | !align=center|[ | + | !align=center|[[Image:Uog.jpg]] || [[Glasgow|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Main Page</font>]] || [[Glasgow/Wetlab|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Wetlab Log</font>]] || [[Glasgow/Modeling|<font face=georgia color=#3366CC size=4>Go To <br> Glasgow's <br>Drylab Log</font>]] |
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| - | |[https://2007.igem.org/Glasgow/Wetlab/Protocols <font face=georgia color=# | + | |[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>] | |[https://2007.igem.org/Glasgow/Wetlab/References <font face=georgia color=#3366CC size=5><b>References</b></font>] | ||
| - | |[https://2007.igem.org/Glasgow/Wetlab/Resources <font face=georgia color=# | + | |[https://2007.igem.org/Glasgow/Wetlab/Resources <font face=georgia color=#3366CC size=5><b>Resources</b></font>] |
|[https://2007.igem.org/Glasgow/Wetlab/Orders <font face=georgia color=#3366CC size=5><b>Orders</b></font>] | |[https://2007.igem.org/Glasgow/Wetlab/Orders <font face=georgia color=#3366CC size=5><b>Orders</b></font>] | ||
| - | |[https://2007.igem.org/Glasgow/Wetlab/Biobricks <font face=georgia color=# | + | |[https://2007.igem.org/Glasgow/Wetlab/Biobricks <font face=georgia color=#3366CC size=5><b>Biobricks<br>Used</b></font>] |
|[https://2007.igem.org/Glasgow/Wetlab/Gels <font face=georgia color=#3366CC size=5><b>Gels</b></font>] | |[https://2007.igem.org/Glasgow/Wetlab/Gels <font face=georgia color=#3366CC size=5><b>Gels</b></font>] | ||
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#Tutorial in [http://www.mbio.ncsu.edu/BioEdit/bioedit.html BioEdit] and [http://biotools.umassmed.edu/bioapps/primer3_www.cgi Primer3]. | #Tutorial in [http://www.mbio.ncsu.edu/BioEdit/bioedit.html BioEdit] and [http://biotools.umassmed.edu/bioapps/primer3_www.cgi Primer3]. | ||
#Started designing primers for amplification and site directed mutagenesis of DmpR and DmpR #24 (see [[Glasgow/Wetlab/Protocols#Protocol 4: Primer Design|Protocol 4]]). | #Started designing primers for amplification and site directed mutagenesis of DmpR and DmpR #24 (see [[Glasgow/Wetlab/Protocols#Protocol 4: Primer Design|Protocol 4]]). | ||
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| + | [[Glasgow/Wetlab/Week2|<font face=georgia color=#3366CC size=4>Next <br> Week</font>]] | ||
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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/Wetlab|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Wetlab Log</font>]] | ||
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Latest revision as of 17:07, 22 October 2007
 | Back To Glasgow's Main Page | Back To Glasgow's Wetlab Log | Go To Glasgow's Drylab Log |
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| Protocols | References | Resources | Orders | Biobricks Used | Gels |
Contents |
Week 1
Tuesday 3rd July 2007
- Maija and Christine prepared LB broth and LB agar with Protocol 1. Made 4 batches of plates, 2 with kanamycin and 2 with carbenicillin.
Wednesday 4th July 2007
- All wetlab researched BioBricks.
- Reporter constructs and mini-Tn5 stocks looked out.
- Streaked the following:
- P. putida PAW 340 pJAK14 (Carb. plate)
- Tn5 lux AB (Carb. plate)
- Mini-Tn5 lux AB (Carb. plate)
- P. fluorescens NCIMB 9815 (Carb. plate)
- P. putida KT2440 (LB)
- JM109 pBluescript 5k+ (Carb. plate)
- Mini-Tn5 Tc (Carb. Plate)
- pQF52 (Carb. plate)
- P. fluorescens 9815 (LB)
- E. coli pJAK14 (Km plate)
- Il DntR in pOF52 (Carb. plate)
- pUCINR in Ω strain C (Carb. plate)
- pGLTUR (Carb. plate)
- Mini-Tn5 Kan (Carb. plate)
- Mini-Tn5 Sm/Sp (Carb. plate)
- Mini-Tn5 1cc2 (Carb. plate)
- E. coli Sa1 (LB)
- DmpR #24 (Carb. plate)
- Mini-Tn5 lac 32 in E. coli 517 (Carb. plate)
- Mini-Tn5 Tc (Carb. Plate)
- Mini-Tn5 Cm (Carb. plate)
- DmpR WT (Carb. plate)
- E.coli sm 10 pESD15 Tn5 GFP (Carb. plate)
- pUJ8 (Carb. Plate)
Thursday 5th July 2007
- BioBricks – Maija and Scott transformed using Protocol 2.
BioBrick Part Function Chassis Plasmid Kit Plate Location (Plate/Well) [http://partsregistry.org/Part:BBa_P1010 Bba_p1010] Construction plasmid E. coli DB3.1 pSB1A10 4/7A " " " pSB1A10 4/11E " " " pSB3K3 4/11C [http://partsregistry.org/Part:BBa_I52001 Bba_I52001] High copy-number construction plasmid E. coli TOP10
should have been DB3.1pSB4A5 4/5I " " " pSB4K5 4/5D " " " pSB4K5 4/6D " " " pSB3K5 4/6B " " " pSB4A5 4/6E [http://partsregistry.org/Part:BBa_J23119 Bba_J23119] Strong constitutive promoter E. coli TOP10 pSB1A2 3/19A [http://partsregistry.org/Part:BBa_R0062 Bba_R0062] HSL & LuxR-inducible promoter E. coli TOP10 pSB1A2 1/9G [http://partsregistry.org/Part:BBa_J04500 Bba_J04500] IPTG-inducible promoter + RBS E. coli TOP10 pSB1AK3 1/16P [http://partsregistry.org/Part:BBa_E0040 Bba_E0040] Promoter-less GFP E. coli TOP10 pSB1A2 1/5H - Maia and Christine researched restriction enzymes for the reporter constructs and Mini-Tn5s streaked on 4/7/07 and digested some. Had planned to make biobricks using Mini-Tn5s etc but discovered that [http://partsregistry.org/Part:BBa_J61206 a previous attempt to use transposable elements to make biobricks] had been unsuccessful due to scarring at the restriction site.
Friday 6th July 2007
- Maija and Christine made 10x stocks for M9 (see Protocol 3.2 - M9).
- Tutorial in [http://www.mbio.ncsu.edu/BioEdit/bioedit.html BioEdit] and [http://biotools.umassmed.edu/bioapps/primer3_www.cgi Primer3].
- Started designing primers for amplification and site directed mutagenesis of DmpR and DmpR #24 (see Protocol 4).
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