MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactreicidal Concentration)

/MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactreicidal Concentration)
MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactreicidal Concentration) 2018-02-23T16:05:44+00:00

MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactreicidal Concentration)

MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactreicidal Concentration)

Introduction

MIC is a method used to determine the lowest concentration of a particular antibiotic needed to kill the bacteria. The microorganisms can be tested for their ability to produce visible growth in microtiter well.

Requirements

  1. Bacterial culture
  2. Nutrient Broth
  3. Sodium Phosphate Buffer
  4. ELISA Reader

Procedure

  1. Inoculate the pathogens in nutrient broth and incubate overnight at 37°C.
  2. Centrifuge the culture at 4000 rpm (1650 G-force) for 10 minutes.
  3. Discard the supernatant and wash the pellet twice with the sodium phosphate buffer.
  4. Measure the absorbance at 610 nm (first the blank: Sodium Phosphate Buffer)
  5. The initial absorbance (A: 0.00) of the suspension was adjusted till 1.00.fill the wells with 100μl Muller Hinton broth except the first well of the column.
  6. Add 100μl plantaricin in the wells A and B.
  7. Dilution starts at well B till well H by transferring 100μl at a time across the column.
  8. Take out 100μl from the well H so that all the wells have a total volume of 100μl.
  9. Add 100μlof pathogen to each well and mix firmly.
  10. Cover the titerwell plate with the aluminum foil at 37°C for 24 hours.
  11. Measure the absorbance of each well with the ELISA reader.
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