Edinburgh/Lab

From 2007.igem.org

(Difference between revisions)
(Biobricks Revived)
(Lab Diary)
Line 40: Line 40:
====Wed 18th July====
====Wed 18th July====
* Digest GFP and mint mini preps and ran on gel
* Digest GFP and mint mini preps and ran on gel
-
* Biobrick [http://partsregistry.org/Part:BBa_E0422 BBa_E0422] (ECFP) revived
+
* Biobrick ECFP BBa_E0422 revived and transformed into E.coli JM109 competent cells
====Thu 19th July====
====Thu 19th July====
* Repeat digest of  GFP and mint mini preps with more DNA
* 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====
====Fri 20th July====
Line 49: Line 50:
* Revived Pseudamonas Fluorescens from freeze dried culture
* Revived Pseudamonas Fluorescens from freeze dried culture
:: Used to retrieve frs and ech genes in vanillin pathway
:: Used to retrieve frs and ech genes in vanillin pathway
-
* Prepare 4 mini preps of ECFP
+
* Made 4 mini preps of ECFP
====Mon 23rd July====
====Mon 23rd July====
* Ran gels for ECFP mini preps
* Ran gels for ECFP mini preps
-
:: Digested using Xba1 and Prt1
+
:: 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

Revision as of 15:40, 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

  • PTG 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 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