Glasgow/Wetlab/Week3
From 2007.igem.org
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|PvuII, PvuI | |PvuII, PvuI | ||
|2250bp, 1030bp, 730bp | |2250bp, 1030bp, 730bp | ||
- | | | + | |- |
- | + | |4/11C | |
- | + | |BBa_p1010 | |
- | + | |pSB3K3 | |
+ | |death gene | ||
+ | |BamHI, XhoI | ||
+ | |190bp, 2390bp,840bp | ||
+ | |- | ||
+ | |1/5H | ||
+ | |BBa_E0040 | ||
+ | |pSB1A2 | ||
+ | |GFP no promoter | ||
+ | |Hime II, PvuI | ||
+ | |1630bp, 1170bp | ||
+ | |- | ||
+ | |4/6D | ||
+ | |BBa_I52001 | ||
+ | | p5B4K5 | ||
+ | |AvaI, PvuI | ||
+ | |920bp, 1470bp, 2120bp | ||
|} | |} | ||
Revision as of 15:32, 17 August 2007
Glasgow Main Page | Back To Wetlab Log
Contents |
Week 3
Tuesday 17th July 2007
- Restriction Digests of transformed BioBricks (see Protocol 7):
Label | BioBrick | Plasmid | Description | Enzymes | Expected Sizes |
---|---|---|---|---|---|
3/19A | BBa_J23119 | pSB1A2 | strong constitutive promoter | NheI, PvuI | 680bp, 1430bp |
1/9G | BBa_R0062 | pBB1A2 | HSL and luxR | EcoRI, PvuI | 1460bp, 660bp |
1/16P | BBa_J04500 | pSB1AK3 | IPTG inducer and RBS | PvuII, PvuI | 2250bp, 1030bp, 730bp |
4/11C | BBa_p1010 | pSB3K3 | death gene | BamHI, XhoI | 190bp, 2390bp,840bp |
1/5H | BBa_E0040 | pSB1A2 | GFP no promoter | Hime II, PvuI | 1630bp, 1170bp |
4/6D | BBa_I52001 | p5B4K5 | AvaI, 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