Team:Heidelberg/Notebook/ViroBytes/October

From 2010.igem.org

(Difference between revisions)
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*~200ng of fragment DNA per reaction
*~200ng of fragment DNA per reaction
*ligation time ~10min
*ligation time ~10min
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==27/09/2010==
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AAV 1 fragments 1-4; 5-8 and 1-8 (full capsid) assembly
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[[Image:Full_assembly_27.10.10.jpg|thumb|600px|right|]]
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**Gel loading scheme:
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**M - GeneRuler 1kb DNA ladder
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**F 1-4 - fragments 1-4 assembly
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**F 5-8 - fragments 5-8 assembly
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**F 1-8 - fragments 1-8 assembly
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**C 1-4 - fragments 1-4 control
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**C 5-8 - fragments 5-8 control
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**C 1-8 - fragments 1-8 control
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{{:Team:Heidelberg/Single_Bottom}}
{{:Team:Heidelberg/Single_Bottom}}
{{:Team:Heidelberg/Single_Bottom}}

Revision as of 03:10, 28 October 2010

October

4/10/2010

  • digest of 8 fragments with BsaI-HF @ 37°C for 1hr
    • 6x2ul of F1-8
    • 3ul NEB4 10x
    • 3ul BSA 10x
    • 11ul H2O
    • 1ul BsaI-HF
  • purification with Nucleotide Removal Kit
  • NanoDrop concentration check:


Fragment 1 Fragment 2 Fragment 3 Fragment 4 Fragment 5 Fragment 6 Fragment 7 Fragment 8
35.5 ng/ul 43.5 ng/ul 43.0 ng/ul 43.0 ng/ul 79.0 ng/ul 51.0 ng/ul 63.0 ng/ul 44.0 ng/ul


5/10/2010

  • assembly started with Quick ligase kit
  • ~200ng of fragment DNA per reaction
  • ligation time ~10min

27/09/2010

AAV 1 fragments 1-4; 5-8 and 1-8 (full capsid) assembly

Full assembly 27.10.10.jpg


    • Gel loading scheme:
    • M - GeneRuler 1kb DNA ladder
    • F 1-4 - fragments 1-4 assembly
    • F 5-8 - fragments 5-8 assembly
    • F 1-8 - fragments 1-8 assembly
    • C 1-4 - fragments 1-4 control
    • C 5-8 - fragments 5-8 control
    • C 1-8 - fragments 1-8 control

Retrieved from "http://2010.igem.org/Team:Heidelberg/Notebook/ViroBytes/October"