Glasgow/Wetlab/Week3
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- | [[Glasgow|Glasgow Main Page]] | + | {| valign=top cellpadding=3 |
+ | |- | ||
+ | !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%" | ||
+ | |- align=center | ||
+ | |[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/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/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>] | ||
+ | |} | ||
---- | ---- | ||
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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 | ||
+ | |High copy death gene | ||
+ | |AvaI, PvuI | ||
+ | |920bp, 1470bp, 2120bp | ||
|} | |} | ||
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#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 | + | #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" | |
- | + | !Template | |
- | + | !Primers | |
+ | |- | ||
+ | |pGLTUR | ||
+ | | | ||
+ | <br>XylR_prefix + XylR_suffix | ||
+ | <br>Pr_prefix + Pr_suffix | ||
+ | <br>Pr_prefix + XylR_suffix | ||
+ | <br>Pu_prefix_EM + Pu_suffix_EM | ||
+ | |- | ||
+ | |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]. | #*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 === | === Thursday 19th July 2007 === | ||
- | #[[User:Mojs|Maia]] redid PCR on pGLTUR and pQF52 using Reddymix and a gradient program ( | + | #[[User:Mojs|Maia]] redid PCR on pGLTUR and pQF52 using Reddymix and a gradient program (see [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|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 | #*XylR prefix and XylR suffix | ||
#*Pr prefix and Pr suffix | #*Pr prefix and Pr suffix | ||
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=== Friday 20th July 2007 === | === Friday 20th July 2007 === | ||
+ | |||
+ | <br> | ||
+ | {| valign=top cellpadding=3 | ||
+ | |- | ||
+ | ![[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>]] | ||
+ | |- | ||
+ | |} | ||
+ | ---- | ||
+ | {| valign=top cellpadding=3 | ||
+ | |- | ||
+ | !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 |
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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
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