IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/08/04: Difference between revisions

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| colspan="2"|
==Team Allergy==
==Team Allergy==
*amiRNA
*amiRNA fragment production appears to have worked at every Tm we tried (click images for details):
[[Image:http://openwetware.org/wiki/Image:AmiRNA-try-3.jpg|240px]][[Image:AmiRNA_pcr_8.3|240px]]
 
*V9/V10  
*V9/V10  
**Diagnostic digest of our gel :
**Diagnostic digest of our gel :
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Gel 1:  Ladder, 9, 11c1, 11c2, ladder, 25c1, 28c1, 28c2, 36c1, 36c2, ladder
Gel 1:  Ladder, 9, 11c1, 11c2, ladder, 25c1, 28c1, 28c2, 36c1, 36c2, ladder
Gel 2: Ladder, 37c1, 37 c2, 38c1, 38c2, ladder
Gel 2: Ladder, 37c1, 37 c2, 38c1, 38c2, ladder


==Team Fence==
==Team Fence==

Revision as of 10:34, 5 August 2010

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Team Allergy

  • amiRNA fragment production appears to have worked at every Tm we tried (click images for details):

File:Http://openwetware.org/wiki/Image:AmiRNA-try-3.jpg

  • V9/V10
    • Diagnostic digest of our gel :

[[Image: |240 px]]

(Should see ~ 7kb band and an 840 band)

    • Realized that we hadn't gel purified our inserts
    • Gel purification of inserts (entire ihpRNA parts)

Gel 1 [[Image: |240px|Gel 2]] Gel 1: Ladder, 9, 11c1, 11c2, ladder, 25c1, 28c1, 28c2, 36c1, 36c2, ladder Gel 2: Ladder, 37c1, 37 c2, 38c1, 38c2, ladder

Team Fence


Team Flavor

Confirmation Digests

  • Ran confirmation digests of miniprepped J45004-1, J45004-2, J45017-1, J45017-2
    • Digested approximately 500ng with xbaI/speI
  • Ran confirmation digests of miniprepped Miraculin N strep+stop and V24, Miraculin C strep+stop and V24, Brazzein N strep+stop and V24, Brazzein C strep+stop and V24.
    • Digested 400-500ng with notI/speI

Retry Ligating j45004 and v0120

  • Used PCR purified J45004 that had been previously digested with xbaI/pstI
  • Used v0120 from Team Vector that they had digested with xbaI/pstI and used in a successful ligation
  • 3:1 insert to backbone ration used in ligation
    • 52ng of insert and 50ng v0120 backbone
    • J45004 insert is 1100 bp
  • Transformed and plated on LB+Amp plates and left overnight


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