What is the purpose of the test? The purpose is to see if the microbe can use the compound malonate as its sole source of carbon and energy for growth.
How is malonate use determined? If a microbe can use malonate for carbon and energy, it will grow on malonate broth . The use of malonate leads to a rise in pH of the medium, and a pH indicator changes color.
What medium is used? The medium used is malonate broth. It contains mineral salts, sodium citrate for carbon, and ammonium phosphate for its nitrogen source. The pH indicator is brom thymol blue, which is green at neutral pH , yellow at acidic pH <6.0 and turns blue at alkaline (basic) pH >7.6.
How is the test performed? An inoculum from a pure culture is transferred aseptically to a sterile tube of malonate broth. The inoculated tube is incubated at 35-37 C for 24 hours and the results are determined. Abundant growth and a change from green to blue in the medium indicates a positive test for growth using malonate .
What reagents are added? None.
To perform this test in VUMIE Online© Online Lab , complete the following steps:
Inoculation of Medium
1. Select the malonate broth medium.
2. Complete the process of a tube-to-tube aseptic transfer to inoculate the medium. Forgotten how to do these things? Watch the "Show Me How To" videos.
Incubation of the Inoculated Medium
3. Place the inoculated tube into the 35-37 C incubator.
4. Press the New Day button to move forward 24 hours. Forgotten how to do these things? Watch the "Show Me How To" videos.
Determination of Test Results
5. Incubate this test for only 24 hours.
6. Retrieve the incubated culture from the incubator.
7 . Observe test result. If the test was followed as described above, the culture will have changed to blue in the presence of alkali/bases (indicating a positive test). In negative tests, there is rarely any growth.
8. Record test result.