Preparation of Stock Solution of Kanamycin (50 mg/ml)

Overview

• Kanamycin is a water-soluble, aminoglycoside bactericidal antibiotic.
• It was originally purified from the bacterium Streptomyces kanamyceticus.
• It inhibits protein synthesis in prokaryotes by binding to the 30S subunit of the bacterial ribosomes.
• Kanamycin is effective against a wide variety of gram-negative and some gram-positive bacteria.

Requirement

Reagents
> Kanamycin
> Deionized / Milli-Q water

Equipment and disposables
> Measuring cylinder
> Conical flask / Beaker / 15 ml screw-capped tube
> Magnetic stirrer

Composition
50 mg/ml kanamycin

Objective
Preparation of 10 ml of 50 mg/ml kanamycin stock solution in water

Preparation

Step 1: To prepare 10 ml of stock solution of kanamycin, weigh out 500 mg of kanamycin. Transfer it to a 15 ml screw-capped tube. Add 9 ml of deionized / Milli-Q water. Mix until all kanamycin is completely dissolved.

Tip:
We recommend using a 15 ml screw-capped tube with milliliter marks. Transfer all the contents in it. Add 9 ml deionized / Milli-Q water and mix the content by rotating the tube.

Precautions:
1. Do not dissolve in 10 ml of deionized / Milli-Q water. It is always a good practice to dissolve all ingredients first and then adjust the volume to the desired volume with the solvent. 
2. Use autoclaved deionized / Milli-Q water.

Step 2: Adjust the volume to 10 ml with deionized / Milli-Q water. Mix it again.

Storage
Store the solution in small aliquots (1 ml) at -20°C for a long time.

Applications
A selection marker for bacterial culture in gene cloning experiments.