UA Biophysics:Protocols:Kanamycin

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Kanamycin disulfate salt from Streptomyces kanamyceticus. Aminoglycoside antibiotic

Kanamycin acts by binding to the 70S ribosomal subunit, inhibiting translocation and eliciting miscoding. Is effective against gram-negative and gram-positive bacteria, and mycoplasma.

STOCK SOLUTION

Kanamycin is usually used at a concentration of 25-50 µg/mL. Stock solution is typically at 25mg/mL

PROTOCOL (STORAGE)

  1. According to desired amount of antibiotics, calculate the amount in grams that has to be weighed.
    1. For example, for 10mL of 25mg/mL of Kanamycin:

25mg/mL*10mL=250mg

  1. Kanamycin is soluble in ddH2O.
  2. Filter sterilize solution using a 0.2 µm syringe filter.
  3. Divide in aliquots of 500μL
  4. Store at -20 °C. up to 4-6 months

It is best to avoid repeated freeze/thaw cycles of the kanamycin aliquots. Kanamycin is light sensitive. Keep tightly closed in light-resistant containers. "Solutions are stable at 37°C for approximately 5 days. Aqueous stock solutions can be stored at 2-8°C for long term storage", Sigma. Plates with LB can be stored one month.

USAGE

For culture plates, allow media to cool to 55 °C before adding ampicillin to a final concentration of 35-50 μg/ml.

For Example, for 300 mL of media at a final concentration of 50µg/mL:


Ci Vi = Cf Vf

25mg/mL* Vi = 0,05mg/mL*300mL

Vi = 15mg/(25mg/mL)

Vi = 0,6 mL

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