Common Buffers, Media, and Stock Solutions


Publication Name:  Current Protocols in Human Genetics
Unit Number:  Appendix 2D
DOI:  10.1002/0471142905.hga02ds26
Online Posting Date:  May, 2001
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Abstract

This appendix describes the preparation of selected bacterial media and of buffers and reagents used in the manipulation of nucleic acids and proteins. Recipes for cell culture media and reagents are located elsewhere in the manual.

    Recipes

  • Acids, concentrated stock solutions

  • Ammonium acetate, 10 M

  • Ammonium hydroxide, concentrated stock solution

  • ATP, 100 mM

  • BCIP, 5% (w/v)

  • BSA (bovine serum albumin), 10% (100 mg/ml)

  • Denhardt solution, 100×

  • dNTPs: dATP, dTTP, dCTP, and dGTP

  • DTT, 1 M

  • EDTA, 0.5 M (pH 8.0)

  • Ethidium bromide solution

  • Formamide loading buffer, 2×

  • Gel loading buffer, 6×

  • HBSS (Hanks balanced salt solution)

  • HCl, 1 M

  • HEPES‐buffered saline, 2×

  • KCl, 1 M

  • LB medium

  • LB plates

  • Loading buffer

  • 2‐ME, (2‐mercaptoethanol)50 mM

  • MgCl2, 1 M

  • MgSO4, 1 M

  • NaCl, 5 M

  • NaOH, 10 M

  • NBT (nitroblue tetrazolium chloride), 5% (w/v)

  • PCR amplification buffer, 10×

  • Phosphate‐buffered saline (PBS), pH ∼7.3

  • Potassium acetate buffer, 0.1 M

  • Potassium phosphate buffer, 0.1 M

  • RNase a stock solution (DNase‐free), 2 mg/ml

  • SDS, 20%

  • SOC medium

  • Sodium acetate, 3 M

  • Sodium acetate buffer, 0.1 M

  • Sodium phosphate buffer, 0.1 M

  • SSC (sodium chloride/sodium citrate), 20×

  • SSPE (sodium chloride/sodium phosphate/EDTA), 20×

  • T4 DNA ligase buffer, 10×

  • TAE buffer, 50×

  • TBE buffer, 10×

  • TBS (Tris‐buffered saline)

  • TCA (trichloroacetic acid), 100% (w/v)

  • TE buffer

  • Terrific broth (TB)

  • Tris⋅Cl, 1 M

  • TY medium, 2×

  • Urea loading buffer, 2×

     
 
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Table of Contents

  • General Guidlines
  • Storage
  • Special Considerations for PCR Experiments
  • Special Considerations for Working with RNA
  • Selection of Buffers
  • Recipes
  • Acknowledgement
  • Tables
     
 
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Materials

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Literature Cited

Literature Cited
   Chemical Rubber Company. 1975. CRC Handbook of Biochemistry and Molecular Biology, Physical and Chemical Data, 3rd ed., Vol. 1. CRC Press, Boca Raton, Fla.
   Mohan, C. (ed.). 1997. Buffers: A Guide for the Preparation and Use of Buffers in Biological Systems. Calbiochem, San Diego, Calif.
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