Haynes Lab:Notebook/ASU iGEM

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::*Colonies observed
::*Colonies observed
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===6-19-12===
 
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*Transformation (LSE)
 
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:*Transformed DNA:
 
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::*T7 promoter BBa_I712074
 
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::*Constitutive promoter BBa_J23102
 
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:* Cells: DH5 alpha (donated)
 
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:* Protocol from: http://tools.invitrogen.com/content/sfs/manuals/subcloningefficiencydh5alpha_man.pdf (30 minute transformation)
 
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:* Controls: puc19, no DNA
 
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:* Plated 2 copies of each (100 ul, 250 ul) on LB amp plates.
 
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* Made 50 LB Amp plates.
 

Revision as of 20:29, 3 July 2012

ASU iGEM 2012

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Project Description/Abstract

Team Chimeric Reporter:

  • Hyder Hussain
  • Abhinav Markus
  • Ryan Muller
  • Ethan Ward

Team Cell Surface Reporter:

  • Nisarg Patel
  • Ryan Muller
  • Rohit Rajan
  • Ellen Qin
  • Melinda Jenner
  • Joe Barth

BioBricks

Magainin Reporter

  • Magainin
  • Magainin + 2xGS Linker + His-Tag
  • Bgal fragment (a-1)
  • Bgal fragment (a-4)
  • Bgal fragment (w)
  • Bgal fragment (1-w)
  • Magainin + 2aa GS Linker + His-Tag + 10aa GS Linker + Bgal fragment (a-1)
  • Magainin + 2aa GS Linker + His-Tag + 10aa GS Linker + Bgal fragment (a-4)
  • Magainin + 2aa GS Linker + His-Tag + 10aa GS Linker + Bgal fragment (w)
  • Magainin + 2aa GS Linker + His-Tag + 10aa GS Linker + Bgal fragment (1-w)
  • His-Tag + 10aa GS Linker + Bgal fragment (a-1)
  • His-Tag + 10aa GS Linker + Bgal fragment (a-4)
  • 13. His-Tag + 10aa GS Linker + Bgal fragment (w)
  • 14. His-Tag + 10aa GS Linker + Bgal fragment (1-w)

Notes

6-15-12

  • Competent cell prep
  • Centrifuged falcon test tubes containing liquid colonies
  • Resuspended in CaCl2 buffer solution and incubated for 15 mins
  • Centrifuged and resuspended in CaCl2 glycerol buffer solution
  • Chilled overnight

6-16-12

  • Competent cell prep
  • Aliquotted 200uL into test tubes
  • Stored in -80C

6-17-12

  • Streak plated prepared competent cells on LB no amp plate
  • Colonies observed




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