IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/09: Difference between revisions

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'''Gel'''
'''Gel'''
[[Image:Adding stop codon.jpg|| 350px]]
    Gel Lanes:
    1. 1kb plus ladder
    3. Miraculin & YFP N-terminus ecoRI/SpeI
    5. Miraculin & YFP C-terminus ecoRI/SpeI
    7. Brazzein & YFP N-terminus ecoRI/SpeI
    9. Brazzein & YFP C-terminus ecoRI/SpeI
    11. V24 NotI/SpeI
    13. NosT & Stop EcoRI/XbaI
 
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Revision as of 13:01, 9 July 2010

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Team Flavor

Ligation of Brazzein & Miraculin (w/ YFP tags) into DUET vector

Digestion

Digestion Reactions
Brazzein & YFP C EcoRI/SpeI Brazzein & YFP N EcoRI/SpeI Miraculin & YFP C EcoRI/SpeI Miraculin & YFP N EcoRI/SpeI V24 NotI/SpeI NosT & Stop EcoRI/XbaI
DNA 2 2 2 3 5 2
FD Buffer (10x) 2 2 2 2 2 2
diH2O 14 14 14 13 11 14
Enzyme 1 1 1 1 1 1
Enzyme 1 1 1 1 1 1
  • We are adding the NosT & stop to the C-terminus of the Miraculin and Brazzein constructs. This whole construct will then be ligated into the V24 DUET vector.

Gel 

   Gel Lanes:
    1. 1kb plus ladder
    3. Miraculin & YFP N-terminus ecoRI/SpeI
    5. Miraculin & YFP C-terminus ecoRI/SpeI 
    7. Brazzein & YFP N-terminus ecoRI/SpeI 
    9. Brazzein & YFP C-terminus ecoRI/SpeI 
   11. V24 NotI/SpeI
   13. NosT & Stop EcoRI/XbaI




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