Glasgow/Wetlab/Week1
From 2007.igem.org
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== Week 1 == | == Week 1 == | ||
| - | === Tuesday | + | === Tuesday 3 July 2007=== |
| - | #Maija and Christine prepared LB broth and LB agar with [[Glasgow/Wetlab/Protocols# Protocol 1: LB Broth and Agar|Protocol 1]]. Made 4 batches of plates, 2 with kanamycin and 2 with carbenicillin. | + | #[[User:MaijaP|Maija]] and [[User:Christinemerrick|Christine]] prepared LB broth and LB agar with [[Glasgow/Wetlab/Protocols# Protocol 1: LB Broth and Agar|Protocol 1]]. Made 4 batches of plates, 2 with kanamycin and 2 with carbenicillin. |
| - | === Wednesday | + | === Wednesday 4 July 2007=== |
#All wetlab researched BioBricks. | #All wetlab researched BioBricks. | ||
#Reporter constructs and mini-Tn5 stocks looked out. | #Reporter constructs and mini-Tn5 stocks looked out. | ||
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#*pUJ8 (Carb. Plate) | #*pUJ8 (Carb. Plate) | ||
| - | === Thursday | + | === Thursday 5 July 2007 === |
| - | #BioBricks – Maija and Scott transformed using [[Glasgow/Wetlab/Protocols#Protocol 2: Transforming Biobricks | Protocol 2]]. | + | #BioBricks – [[User:MaijaP|Maija]] and [[User:scott.w.ramsay|Scott]] transformed using [[Glasgow/Wetlab/Protocols#Protocol 2: Transforming Biobricks | Protocol 2]]. |
#*BBa_p1010 (DB3.1) (death gene plasmid) Plate 4: 7A – p5B1A10, 11E – p5B1A10 and 11C – p5B3K3 | #*BBa_p1010 (DB3.1) (death gene plasmid) Plate 4: 7A – p5B1A10, 11E – p5B1A10 and 11C – p5B3K3 | ||
#*BBa_IS2001 (Top10) (high copy number plasmid) Plate 4: 5I – p5B4A5, 5D – p5B4K5, 6B – p5B3K5, 6D – p5B4K5 and 6E – p5B4A5 (Also 5K, 5M, 4J, 4L, 4N, 4P) | #*BBa_IS2001 (Top10) (high copy number plasmid) Plate 4: 5I – p5B4A5, 5D – p5B4K5, 6B – p5B3K5, 6D – p5B4K5 and 6E – p5B4A5 (Also 5K, 5M, 4J, 4L, 4N, 4P) | ||
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#*BBa_J04500 (Top 10) (IPTG inducible prom + RBS) Plate 1: 16P – p5B1AK3 (V1009) | #*BBa_J04500 (Top 10) (IPTG inducible prom + RBS) Plate 1: 16P – p5B1AK3 (V1009) | ||
#*BBa_E0040 (Top 10) (GFP mutant no promoter (3b)) Plate 1: 5H – p5B1A2 (V1001) | #*BBa_E0040 (Top 10) (GFP mutant no promoter (3b)) Plate 1: 5H – p5B1A2 (V1001) | ||
| - | #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 a previous attempt to use transposable elements to make biobricks had been unsuccessful due to scarring at the restriction site. http://partsregistry.org/Part:BBa_J61206 | + | #[[User:Mojs|Maia]] and [[User:Christinemerrick|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 a previous attempt to use transposable elements to make biobricks had been unsuccessful due to scarring at the restriction site. http://partsregistry.org/Part:BBa_J61206 |
| - | === Friday | + | === Friday 6 July 2007 === |
| - | #Maija and Christine made 10x stocks for M9 (see [[Glasgow/Wetlab/Protocols#Protocol 3: Minimal Media and Trace Elements|Protocol 3.2 - M9]]). | + | #[[User:MaijaP|Maija]] and [[User:Christinemerrick|Christine]] made 10x stocks for M9 (see [[Glasgow/Wetlab/Protocols#Protocol 3: Minimal Media and Trace Elements|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]. | #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]]). | ||
Revision as of 18:27, 15 August 2007
Contents |
Week 1
Tuesday 3 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 4 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 5 July 2007
- BioBricks – Maija and Scott transformed using Protocol 2.
- BBa_p1010 (DB3.1) (death gene plasmid) Plate 4: 7A – p5B1A10, 11E – p5B1A10 and 11C – p5B3K3
- BBa_IS2001 (Top10) (high copy number plasmid) Plate 4: 5I – p5B4A5, 5D – p5B4K5, 6B – p5B3K5, 6D – p5B4K5 and 6E – p5B4A5 (Also 5K, 5M, 4J, 4L, 4N, 4P)
- BBa_J23119 (Top 10) (strong constitutive promoter) Plate 3: 19A – pB1A2 (V1013)
- BBa_R0062 (Top 10) (HSL and luxR inducible) Plate 1: 9G – pSB1A2 (V1004)
- BBa_J04500 (Top 10) (IPTG inducible prom + RBS) Plate 1: 16P – p5B1AK3 (V1009)
- BBa_E0040 (Top 10) (GFP mutant no promoter (3b)) Plate 1: 5H – p5B1A2 (V1001)
- 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 a previous attempt to use transposable elements to make biobricks had been unsuccessful due to scarring at the restriction site. http://partsregistry.org/Part:BBa_J61206
Friday 6 July 2007
- Maija and Christine made 10x stocks for M9 (see Protocol 3.2 - M9).
- Tutorial in BioEdit and Primer3.
- Started designing primers for amplification and site directed mutagenesis of DmpR and DmpR #24 (see Protocol 4).
