Haynes:Protocols
Protocols
BioBrick Assembly
- Assembly of BioBrick Parts: an Overview
- Making Standardized DNA Parts
- Model Procedure for Assembling Parts: Classic Ligation for Beginners
Cell Culture: Bacteria
- Chemically competent cell prep
- Glycerol Stocks (long term -80°C storage)
Cell Culture: Mammalian
- Coming soon
Real Time Quantitative PCR
- Universal Probe Library (UPL) assay: for the Roche Light Cycler 480
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
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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.