Glasgow/Wetlab/Week3
From 2007.igem.org
| (2 intermediate revisions not shown) | |||
| Line 1: | Line 1: | ||
{| valign=top cellpadding=3 | {| valign=top cellpadding=3 | ||
|- | |- | ||
| - | !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>]] |
|} | |} | ||
---- | ---- | ||
{|cellspacing="6px" cellpadding="16" border="0" width="100%" | {|cellspacing="6px" cellpadding="16" border="0" width="100%" | ||
|- align=center | |- align=center | ||
| - | |[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>] | ||
|} | |} | ||
| 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" | ||
| Line 106: | Line 106: | ||
{| valign=top cellpadding=3 | {| valign=top cellpadding=3 | ||
|- | |- | ||
| - | ![[Glasgow/Wetlab/Week2|<font face=georgia color=# | + | ![[Glasgow/Wetlab/Week2|<font face=georgia color=#3366CC size=4>Previous <br> Week</font>]] || [[Glasgow/Wetlab/Week4|<font face=georgia color=#3366CC size=4>Next <br> Week</font>]] |
|- | |- | ||
|} | |} | ||
| Line 112: | Line 112: | ||
{| valign=top cellpadding=3 | {| valign=top cellpadding=3 | ||
|- | |- | ||
| - | !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>]] |
|} | |} | ||
Latest revision as of 17:08, 22 October 2007
 | 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 |
|---|
| | Back To Glasgow's Main Page | Back To Glasgow's Wetlab Log |
|---|
