IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/06/15: Difference between revisions

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Used RNeasy Plant Mini Kit to extract RNA from the flavedo of an Organic Valencia Orange.
Used RNeasy Plant Mini Kit to extract RNA from the flavedo of an Organic Valencia Orange.


[[Image:Orange_cross_section_description.png]]
[[Image:Flavedo.png]]




Revision as of 14:17, 15 June 2010

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Team Flavour

Primer Designs for Agrobacterium Vector

  • Primers were designed to amplify from the Expression Series pORE Agrobacterium plasmid (e3) 1)the pENTcup2 promoter, 2) the tNOS stop sequence, 3) the tNOS stop sequence + additional stop codon and 4) the pHLP promoter.

Note: the NOSterm_BB_R primer works for both tNOS and tNOS+STOP codon sequences. 

   pENTcup2_BB_F
   CCTTTCTAGAGGGATCTTCTGCAAGCATCT

   pENTcup2_BB_R
   AAGGCTGCAGCGGCCGCTACTAGTTCCGGTGGGTTTTGAGGT

   STOP_NOSterm_BB_F
   CCTTTCTAGATGAGATCGTTCAAACATTTGG

   NOSterm_BB_F
   CCTTTCTAGAGATCGTTCAAACATTTGGCA

   NOSterm_BB_R
   AAGGCTGCAGCGGCCGCTACTAGTGATCTAGTAACATAGATGACA

   pHPL_BB_F
   CCTTTCTAGAAACGTGGATACTTGGCAGTG

   pHPL_BB_R
   AAGGCTGCAGCGGCCGCTACTAGTCTTTTGAGCTTAGAGGTTTTT

BioBrick Transformation

  • Colonies were observed after overnight culture growth, but in low quantities.
    • J45700 and J45004 (both Wintergreen Pathway) showed minimal number of colonies on both 10μL and 100μL cultures.
    • J45250 and J45014 (both Banana Pathway) showed no colonies on either the 10μL or 100μL cultures.

Codon Usage in Arabidopsis

Using http://gcua.schoedl.de/

Wintergreen Pathway:
BioBrick Part #: BBa_J45700
Original Organism: Pseudomonas aeruginosa

BBa_J45700 BBa_J45700 BBa_J45700 BBa_J45700

Banana Pathway:
BioBrick Part #: BBa_J45014
Original Organism: Saccharomyces cerevisiae
BBa_J45014 BBa_J45014 BBa_J45014

BioBrick Part #: BBa_J45250
BBa_J45250 BBa_J45250 BBa_J45250 BBa_J45250

Valencene Pathway:

Valencene Valencene Valencene

Valencene Extraction

Used RNeasy Plant Mini Kit to extract RNA from the flavedo of an Organic Valencia Orange.



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