Recipe: Alkaline Methylene Blue 0.01%

Alkaline methylene blue solution – 100 ml

Chemicals:

• Methylene blue (Sigma)
• 0.1M Glycine buffer (pH 10.6)
• Distilled water

Procedure:

• Dissolve 0.1g methylene blue in 100 ml distilled water (0.1% MB stock solution).
• Mix 10 ml of the MB stock solution with 90 ml of the 0.1M glycine buffer. This solution is used for the viability staining.

0.1M Glycine buffer (pH 10.6) – 100 ml

Chemicals:

• 1M NaOH solution (Merck)
• Glycine (Sigma)
• Distilled water

Procedure:

• Dissolve 0.75 g glycine in 70 ml water.
• Adjust the pH with the NaOH solution to 10.6.
• Fill volume to 100 ml with distilled water