Glasgow/Wetlab/Week3
From 2007.igem.org
(Difference between revisions)
(→Wednesday 18th July 2007) |
|||
| Line 72: | Line 72: | ||
#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). | #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). | ||
#[[User:MaijaP|Maija]] ran a gel of the restriction digests to check the sizes of the biobricks. These were the maps we made yesterday. | #[[User:MaijaP|Maija]] ran a gel of the restriction digests to check the sizes of the biobricks. These were the maps we made yesterday. | ||
| - | #[[User:Mojs|Maia]] is extracting DNA from 3 samples of | + | #[[User:Mojs|Maia]] is extracting DNA from 3 samples of Pseudomonas Aeruginosa, which were plated yesterday and grown in liquid media, using MoBio PowersoilTM DNA purification kit (see [[Glasgow/Wetlab/Protocols#Protocol 8: MoBio PowersoilTM DNA Purification Kit|Protocol 8]]); only change was to shorten "shaking" time from 10 to 2 minutes. |
#PCR trial run using Reddymix and Touch 2 done (see [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|Protocol 9]] for PCR). | #PCR trial run using Reddymix and Touch 2 done (see [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|Protocol 9]] for PCR). | ||
{| border="1" cellspacing="0" cellpadding="5" align="center" | {| border="1" cellspacing="0" cellpadding="5" align="center" | ||
Revision as of 15:13, 19 October 2007
| https://static.igem.org/mediawiki/2007/thumb/c/cc/Uog.jpg/50px-Uog.jpg | Back To Glasgow's Main Page | Back To Glasgow's Wetlab Log | Go To Glasgow's Drylab Log |
|---|
| Protocols | References | Resources | Orders | Biobricks Used | Gels |
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 | High copy death gene | 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 Pseudomonas Aeruginosa, 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 (see Protocol 9 for PCR).
| Template | Primers |
|---|---|
| pGLTUR |
|
| pQF52 | 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].
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
Friday 20th July 2007
| Previous Week | Next Week |
|---|
| https://static.igem.org/mediawiki/2007/thumb/c/cc/Uog.jpg/50px-Uog.jpg | Back To Glasgow's Main Page | Back To Glasgow's Wetlab Log |
|---|
