Haynes Lab:Notebook/ASU iGEM

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:* Protocol from: http://tools.invitrogen.com/content/sfs/manuals/subcloningefficiencydh5alpha_man.pdf (30 minute transformation)
:* Protocol from: http://tools.invitrogen.com/content/sfs/manuals/subcloningefficiencydh5alpha_man.pdf (30 minute transformation)
:* Controls: puc19, no DNA
:* Controls: puc19, no DNA
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===6-21-12===
 
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*Made Liquid Cultures of E.coli transformed with RBS B0030
 
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*Made Liquid Cultures of E.coli transformed with TetR GFP
 
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*miniprepped and nanodropped T7 promoter BBa_I712074 and Constitutive promoter BBa_J23102 liquid cultures
 
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* liquid cultures:
 
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:* RBS1
 
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:* RBS2 (duplicate_
 
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:* GFP1
 
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:* puc19
 
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:* negative controls
 
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:* 5 ml LB amp
 
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:* overnight cultures
 
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* replated GFP1 & 2 (duplicates)
 
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*Nanodropped plasmid DNA samples
 
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:*Constitutive promoter 1: __ng/uL
 
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:*Constitutive promoter 2: __ng/uL
 
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:*T7 promoter 1: __ng/uL
 
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:*T7 promoter 2: __ng/uL
 
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Revision as of 20:24, 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

6-19-12

  • Transformation (LSE)
  • Transformed DNA:
  • T7 promoter BBa_I712074
  • Constitutive promoter BBa_J23102
  • Made 50 LB Amp plates.

6-20-12

  • Plated negative control on LB Amp plate
  • Liquid cultures of T7 promoter and constitutive promoter
  • Transformation (LSE)
  • Transformed DNA:
  • RBS (well 1:1H BBa_B0030)
  • TetR GFP (well 2:8A Part:BBa_I13522)




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