Edinburgh/Lab
From 2007.igem.org
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* designed primers for Sam5 and Sam8 genes (part of vanillin biosynthesis pathway) | * designed primers for Sam5 and Sam8 genes (part of vanillin biosynthesis pathway) | ||
| - | ==== | + | ====Thu 26th July==== |
* Prepared glycerol stock of ''Pseudomonas fluorescens'' | * Prepared glycerol stock of ''Pseudomonas fluorescens'' | ||
* Prepared miniprep of GFP and mint biobricks | * Prepared miniprep of GFP and mint biobricks | ||
* Streaked ''Pseudomonas fluorescens'' onto a fresh LB plate | * Streaked ''Pseudomonas fluorescens'' onto a fresh LB plate | ||
* Revived [http://partsregistry.org/Part:BBa_I0500 BBa_I0500] and [http://partsregistry.org/Part:BBa_Q04510 BBa_Q04510] | * Revived [http://partsregistry.org/Part:BBa_I0500 BBa_I0500] and [http://partsregistry.org/Part:BBa_Q04510 BBa_Q04510] | ||
| + | |||
| + | ====Fri 27th July==== | ||
| + | * Continued miniprep of GFP and mint biobricks | ||
| + | :: Restriction digest using SpeI & EccRI restriction enzymes | ||
| + | * PCR of ''Pseudomonas fluorescens'' genes | ||
Revision as of 10:10, 30 July 2007
https://static.igem.org/mediawiki/2007/f/f5/800px-Edinburgh_City_15_mod.JPG
Contents |
Biobricks Revived
This is a list of the biobricks we have revived from the registry
| Brick | Description | Well | Used for |
|---|---|---|---|
| [http://partsregistry.org/Part:BBa_J04430 BBa_J04430] | IPTG induced GFP | Plate 2, 1L | Test biobrick revival process |
| [http://partsregistry.org/Part:BBa_E0241 BBa_E0241] | PoPS --> GFP | Plate 2, 15L | For use in dif experement 1 |
| [http://partsregistry.org/Part:BBa_J45700 BBa_J45700] | Wintergreen Scent | Plate 3, 7B | Place into lacto bacillus |
| [http://partsregistry.org/Part:BBa_E0422 BBa_E0422] | fast degrading ECFP | Plate 1, 11G | For use in dif experement 1 |
| [http://partsregistry.org/Part:BBa_I0500 BBa_I0500] | arabinose promoter | Plate 2, 9I | For use in dif experement 3 |
| [http://partsregistry.org/Part:BBa_Q04510 BBa_Q04510] | CI (Lambda) Inverter | Plate 2, 13K | For use in dif experement 3 |
Lab Diary
Thu 12th July
- IPTG induced GFP revived ([http://partsregistry.org/Part:BBa_J04430 BBa_J04430])
- Test biobrick revival process
Fri 13th July
- Alfalfa Seeds Planted (in Agar)
- Growing alfalfa plants, to get leaves from which to extract DNA for the caffeic acid-3-o-methyltransferase gene (part of the vanillin synthesis pathway)
- Biobricks [http://partsregistry.org/Part:BBa_E0241 BBa_E0241] and [http://partsregistry.org/Part:BBa_J45700 BBa_J45700] revived, using modified protocal from open wetware site
Mon 16th July
- Begin DNA extraction from Bamboo
- Test DNA extraction process from plants
- Prepared mini preps for GFP and mint genes
Tue 17th July
- Alfalfa Transferred to compost
- DNA containing GFP and mint genes extracted from E.Coli
Wed 18th July
- Double digest GFP and mint mini preps, using restriction enzymes SpeI and EcoRI. These were then run on a polyacramide gel
- Biobrick ECFP [http://partsregistry.org/Part:BBa_E0422 BBa_E0422] revived and transformed into E.coli JM109 competent cells
Thu 19th July
- Repeat of double digest (using SpeI & EcoRI) of GFP and mint mini preps with increased concentration of DNA
- Replate colonies of ECFP, 4 overnight cultures prepared for making DNA mini preps
- designed primers for COMT gene (part of vanillin biosynthesis pathway)
Fri 20th July
- Prepared GFP and winter green biobricks for sequencing
- Revived Pseudamonas fluorescens from freeze dried culture (P. fluorescens contains two genes, ech and fcs, which are part of the synthetic vanillin biosynthesis pathway)
- Made 4 mini preps of ECFP
- Designed primers for ech and fcs genes
- Ordered Saccharothrix espeanensis from DSMZ culture collection, which contains Sam5 and Sam8 genes required for the synthetic vanillin biosynthesis pathway
Mon 23rd July
- Ran gels for ECFP mini preps
- Restriction digested using XbaI and PstI
- Alfalfa growing well - now have enough leaf material from which to extract DNA
Tue 24th July
- Annealed dif F & R oligoes, restriction digested with SacI & SpeI, ligated with BioBrick vector Edinbrick 1
Wed 25th July
- Transformed dif F & R DNA into E.coli, incubated at 37C overnight
- designed primers for Sam5 and Sam8 genes (part of vanillin biosynthesis pathway)
Thu 26th July
- Prepared glycerol stock of Pseudomonas fluorescens
- Prepared miniprep of GFP and mint biobricks
- Streaked Pseudomonas fluorescens onto a fresh LB plate
- Revived [http://partsregistry.org/Part:BBa_I0500 BBa_I0500] and [http://partsregistry.org/Part:BBa_Q04510 BBa_Q04510]
Fri 27th July
- Continued miniprep of GFP and mint biobricks
- Restriction digest using SpeI & EccRI restriction enzymes
- PCR of Pseudomonas fluorescens genes
