Edinburgh/Lab

From 2007.igem.org

(Difference between revisions)
(Thur 26th July)
(Lab Diary)
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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)
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====Thur 26th July====
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====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]
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====Fri 27th July====
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* Continued miniprep of GFP and mint biobricks
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:: Restriction digest using SpeI & EccRI restriction enzymes
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* 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