Edinburgh/Lab

From 2007.igem.org

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* PTG induced GFP revived ([http://partsregistry.org/Part:BBa_J04430 BBa_J04430])
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* IPTG induced GFP revived ([http://partsregistry.org/Part:BBa_J04430 BBa_J04430])
::Test biobrick revival process
::Test biobrick revival process

Revision as of 16:03, 25 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

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

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

  • Digest GFP and mint mini preps and ran on gel
  • Biobrick ECFP [http://partsregistry.org/Part:BBa_E0422 BBa_E0422] revived and transformed into E.coli JM109 competent cells

Thu 19th July

  • Repeat digest of GFP and mint mini preps with more DNA
  • Replate colonies of ECFP, 4 overnight cultures prepared for making DNA mini preps

Fri 20th July

  • Prepared GFP and winter green biobricks for sequencing
  • Revived Pseudamonas Fluorescens from freeze dried culture
Used to retrieve frs and ech genes in vanillin pathway
  • Made 4 mini preps of ECFP

Mon 23rd July

  • Ran gels for ECFP mini preps
Restriction digested using XbaI and PstI


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