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

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''Digestion''
''Digestion''


{| border="1"
|+ ''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
| align="center" | 2 || align="center" |  2|| align="center" | 2|| align="center" | 3|| align="center" | 5||align="center"| 2
|-
! FD Buffer (10x)
| align="center"  | 2 || align="center" |  2 || align="center" |  2|| align="center" |  2|| align="center" |  2|| align="center" |2 
|-
! diH<sub>2</sub>O
| align="center"  | 14 || align="center" |  14 || align="center" |  14|| align="center"  | 13|| align="center"  | 11|| align="center"| 14
|-
! Enzyme
| align="center"  | 1 || align="center" |  1 || align="center" |  1|| align="center" |  1|| align="center" |  1|| align = "center"| 1 
|-
! Enzyme
| align="center" | 1 || align="center" |  1|| align="center" | 1|| align="center" | 1|| align="center" | 1|| align="center" |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.




Revision as of 12:49, 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.



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