IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/08/05

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(Team Allergy)
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==Team Allergy==
==Team Allergy==
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* Insert your content here.
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* Gel extracte V9/V10 backbone
[[Image:V9v10.jpg|240px]]
[[Image:V9v10.jpg|240px]]
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''Concentrations: V9 (9.4 ng/uL; V10 (16.4 ng/uL)
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*Ligated ihpRNA inserts into V9/V10 and transformed
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**For our ligations we only used ~ 2uL of backbone (around 18 and 32 ng of backbone)and used a 3x excess of insert
* Verified that amiRNA stitching of Bet, LTP yielded the proper insert with a low level of background through PCR:
* Verified that amiRNA stitching of Bet, LTP yielded the proper insert with a low level of background through PCR:
[[Image:AmiRNA-insert-diagnostic.jpg|240px]]
[[Image:AmiRNA-insert-diagnostic.jpg|240px]]

Revision as of 11:20, 6 August 2010

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Team Flavor

Transformation of J45004

  • 5μL of registry DNA was transformed with 15μL TURBO cells; mixture was plated on LB+AMP and left to grow O/N

Team Allergy

  • Gel extracte V9/V10 backbone

Concentrations: V9 (9.4 ng/uL; V10 (16.4 ng/uL)

  • Ligated ihpRNA inserts into V9/V10 and transformed
    • For our ligations we only used ~ 2uL of backbone (around 18 and 32 ng of backbone)and used a 3x excess of insert
  • Verified that amiRNA stitching of Bet, LTP yielded the proper insert with a low level of background through PCR:

  • Digested Bet,LTP inserts with X+P, B21 with X+P+phosphatase
  • Ligated, transformed
    • 0-7: Bet, corresponding to 65..55 degrees C for stitching Tm (even spacing)
    • 8-15: LTP, corresponding to 65.55 degrees C during stitching annealing, even spacing



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