50 mg/mL Kanamycin stock

WFID: 10093   Date: 06-18-2011  
Protocol Format: Public Draft.
Evan Brower  
Biochemical, Molecular Biology, Formulation
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Introduction

When cultivating bacterial cultures for the purposes of recombinant protein expression, antibiotics are necessary to provide selection pressure for the expression vector harbored by the bacteria.  Many commercially available vectors provide kanamycin resistance.  Kanamycin is usually used at a concentration of 25-50 µg/mL.  Many people prefer kanamycin over ampillicin due to its greater long-term stability.

Materials

Catalog # Material Name Company Name Solute/Solvent
1K1377Kanamycin sulfateSigmaSolute
243121828mm Diameter Syringe Filters, 0.2Ám Pore SFCA-PF MembraneCorning

Protocol

For preparation of 10 mL of 50 mg/mL kanamycin
1) Dissolve 500 mg kanamycin sulfate powder in 10 mL ddH2O.


2) Filter sterilize antibiotic solution using a 0.2 µm syringe filter.


3) Aliquot (500 µL - 1 mL aliquots are useful) and store at -20 °C until use.


NOTE: It is best to avoid repeated freeze/thaw cycles of the kanamycin aliquots.

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