Haynes:Protocols

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=Protocols=
=Protocols=
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'''BioBrick Assembly'''
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==DNA Assembly==
* [[Haynes:BioBrick Method short | Assembly of BioBrick Parts: an Overview]]
* [[Haynes:BioBrick Method short | Assembly of BioBrick Parts: an Overview]]
* [[Haynes:Making BioBricks | Making Standardized DNA Parts]]
* [[Haynes:Making BioBricks | Making Standardized DNA Parts]]
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'''Cell Culture: Bacteria'''
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==Cell Culture: Bacteria==
* [[Haynes:ChemComp cells | Chemically competent cell prep]]
* [[Haynes:ChemComp cells | Chemically competent cell prep]]
* [[Haynes:Glycerol Stocks | Glycerol Stocks]] - for long term -80°C storage
* [[Haynes:Glycerol Stocks | Glycerol Stocks]] - for long term -80°C storage
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'''Cell Culture: Mammalian'''
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==Cell Culture: Mammalian==
* [[Haynes:MamCultureMedia | Media formulas]] - cell line-specific formulas
* [[Haynes:MamCultureMedia | Media formulas]] - cell line-specific formulas
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'''Protein'''
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==Protein==
* [[Haynes:Bradford | Bradford assay]] - measure protein concentration
* [[Haynes:Bradford | Bradford assay]] - measure protein concentration
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'''Real Time Quantitative PCR'''
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==Real Time Quantitative PCR==
* [[Haynes:UPLassay | Universal Probe Library (UPL) assay]]: for the Roche Light Cycler 480
* [[Haynes:UPLassay | Universal Probe Library (UPL) assay]]: for the Roche Light Cycler 480
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'''Resources at OpenWetWare'''
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==Resources at OpenWetWare==
* [http://openwetware.org/wiki/E._coli_genotypes E. coli Strains]  
* [http://openwetware.org/wiki/E._coli_genotypes E. coli Strains]  
* [http://openwetware.org/wiki/Materials Materials]: buffer formulas; specific information on commonly used reagents (''e.g.'', LB broth, ampicillin, restriction enzymes, etc.)
* [http://openwetware.org/wiki/Materials Materials]: buffer formulas; specific information on commonly used reagents (''e.g.'', LB broth, ampicillin, restriction enzymes, etc.)
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'''Software Guides'''
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==Software Guides==
* [[Haynes:Jmol_Guide | Jmol commands: controlling PDB structure displays]]
* [[Haynes:Jmol_Guide | Jmol commands: controlling PDB structure displays]]

Revision as of 18:36, 27 October 2012

link=Haynes_Lab
link=http://engineering.asu.edu/

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Contents

Protocols

DNA Assembly


Cell Culture: Bacteria


Cell Culture: Mammalian


Protein


Real Time Quantitative PCR


Resources at OpenWetWare

  • E. coli Strains
  • Materials: buffer formulas; specific information on commonly used reagents (e.g., LB broth, ampicillin, restriction enzymes, etc.)


Software Guides



Recipes

Click "show" to expand each recipe, and "hide" to, well, hide it.

Bromo-Blue/X-cyanol Loading Buffer, 10X

Formula: [30% glycerol; 12.5 mg/mL bromophenol blue; 12.5 mg/mL xylene cyanol]
Volume: 20 mL

  • 50% glycerol, 12 mL
  • Bromophenol blue, 250 mg
  • Xylene cyanol, 250 mg

Bring up to total volume with dH2O.

Chromatin Prep Buffer A

Formula: [10 mM HEPES (pH 7.9); 10 mM KCl, 1.5 mM MgCl2; 0.34 M sucrose; 10% glycerol; 1 mM dithiothreitol; 1x protease inhibitor cocktail]
Volume: 100 mL

  • 1 M HEPES pH 7.9, 1 mL
  • 1 M KCl, 1 mL
  • 1 M MgCl2, 150 μL
  • Sucrose, 11.6 g
  • 50% Glycerol, 20 mL
  • 1 M Dithiotreitol (DTT), 100 μL

Bring up to total volume with dH2O. Store at 4°C. Add 100x protease inhibitor cocktail* (1:100) immediately before use.

LB Broth (Lennox)

Volume: 1 L

  • Caesin tryptone, 10 g
  • Yeast extract, 5 g
  • NaCl, 5 g
  • 1 N NaOH, 1 mL

Dissolve in 1000 mL dH2O. Autoclave to sterilize. Cool to room temperature before adding antibiotics. Shortcut: Use 25 g Acros LB Broth, Lennox (BP9722-500) granules instead of caesin tryptone, yeast extract, and NaCl.

Mammalian Cell Media

Volume: 500 mL

Add FBS and pen-strep directly to the incomplete medium in the original stock bottle. Cap and invert to mix. Filter sterilize using a vacuum filtration unit.


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