Glasgow/Wetlab/Week3
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#*pQF52 plasmid: DntR_prefix / DntR_suffix | #*pQF52 plasmid: DntR_prefix / DntR_suffix | ||
#*DmpR WT/24: DntR_prefix / DntR_suffix | #*DmpR WT/24: DntR_prefix / DntR_suffix | ||
| - | #*Biobricks: [http://partsregistry.org/Part:BBa_R0062 R0062], [http://partsregistry.org/Part:BBa_E0040 E0040], [http://partsregistry.org/Part:BBa_J04500 J04500], [http://partsregistry.org/Part:BBa_J23119 J23119], http://partsregistry.org/Part:BBa_p1010 p1010], http://partsregistry.org/Part:BBa_I52001 I52001]. | + | #*Biobricks: [http://partsregistry.org/Part:BBa_R0062 R0062], [http://partsregistry.org/Part:BBa_E0040 E0040], [http://partsregistry.org/Part:BBa_J04500 J04500], [http://partsregistry.org/Part:BBa_J23119 J23119], [http://partsregistry.org/Part:BBa_p1010 p1010], [http://partsregistry.org/Part:BBa_I52001 I52001]. |
#*See [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|Protocol 9]] for PCR. | #*See [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|Protocol 9]] for PCR. | ||
Revision as of 18:33, 15 August 2007
Glasgow Main Page | Back To Wetlab Log
Contents |
Week 3
Tuesday 17th July 2007
- Restriction Digests (see Protocol 7):
- BBa_J23119 (strong constitutive promoter) pSB1A2. 1 x NheI, 1 x PvuI. 680bp, 1430bp
- BBa_R0062 (HSL and luxR) pBB1A2. 1 x EcoRI, 1 x PvuI. 1460bp, 660bp
- BBa_J04500 (IPTG inducer and RBS) pSB1AK3. 1 x PvuII, 2 x PvuI. 2250bp, 1030bp, 730bp
- BBa_p1010 (death gene) pSB3K3. 1 x BamHI, 2 x XhoI. 190bp, 2390bp,840bp.
- BBa_E0040 (GFP no promoter) pSB1A2. 1 x Hime II, 1 x PvuI. 1630bp, 1170bp.
- BBa_I52001 (6D) p5B4K5. 2 x AvaI, 1 x PvuI. 920bp, 1470bp, 2120bp.
Wednesday 18th July 2007
- Last night we diluted primers to 100 pmol/ul by adding the amount of dH2O given on the order sheet, and made working solutions to 10 pmol/ul (2 x 200 ul each - 20 ul stock and 180 ul dH2O).
- Maija ran a gel of the restriction digests to check the sizes of the biobricks. These were the maps we made yesterday.
- Maia is extracting DNA from 3 samples of ****, which were plated yesterday and grown in liquid media, using MoBio PowersoilTM DNA purification kit (see Protocol 8); only change was to shorten "shaking" time from 10 to 2 minutes.
- PCR trial run using Reddymix and Touch 2 done on:
Thursday 19th July 2007
- Maia redid PCR on pGLTUR and pQF52 using Reddymix and a gradient program (See Protocol 9) to minimise the non-specific amplification evident in previous gels. New DntR suffix primer has arrived. Used the following primer sets.
- XylR prefix and XylR suffix
- Pr prefix and Pr suffix
- Pr prefix and XylR suffix
- Pu prefix and Pu suffix
- DntR prefix and DntR suffix 2
