IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/19: Difference between revisions

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*Second digest of B21 appears to have been more successful:
*Second digest of B21 appears to have been more successful:


[[Image:Backbone_digest_confirmatio.jpg|200px]]
[[Image:Backbone_digest_confirmatio.jpg|200px]]


*Ligated and transformed
*Ligated and transformed

Revision as of 08:03, 20 July 2010

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Team Flavor

DTL of Mira/Brazz+StrepII into STOP+V0120

Digestion

Digestion Reactions
Stop v0120 1 Stop v0120 2
DNA 10 8
FD Buffer (10x) 2 2
diH2O 6 8
Xba1 1 1
EcoRI 1 1
Ligation Reactions
Mira N + Stop Mira C + Stop Brazz N + Stop Brazz C + Stop Stop only Control
diH2O 3 3 11 6 14
T4 DNA ligase buffer (10x) 2 3 2 2 2
DNA Insert 11 11 3 8 0
DNA Backbone 3 3 3 3 3
T4 DNA ligase 1 1 1 1 1


Confirm of valencene PCR (failed)

  • Re-do of Valencene PCR (failed)

Team Allergy

  • Got back sequencing results of GerS, BetS, PDK
    • All three samples of GerS sent in worked and PDK worked
  • Ran PCR of Friday's digested B21. The gel suggested a partial digest: we saw two bands of backbone where we should have seen only one:

  • Sending 6 samples of amiRNA (LTP.5), 5 samples of amiRNA (LTP), 6 samples of amiRNA (Bet)
  • Second digest of B21 appears to have been more successful:

  • Ligated and transformed
    • allergen parts
    • Pme/pal



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