|just an example|
After MIC, you can proceed to MBC (Minimal Bactericidal Inhibition) and MFC (Minimal Fungicidal Concentration).
For me, the tricky part for this experiment is started when we observe 96-wells plate during MIC to determine which well should be used to proceed to this test. So what I'm doing is observed the turbidity in each well. Clear well determined that the sample is able to inhibit microorganism (bacteria or fungi) at this concentration.
1. Observe 96-wells plate from MIC and determine the clear well.
2. Pipette 100 micro-lite from the clear well into agar and spread.
3. Pipette 100 micro-lite from a well above the clear well into another agar plate. Repeat it for another two times. E.g. : from well H until E. Sometimes, it only require three wells e.g well C until A.
4. Incubate 18 hours for MBC and 72 hours for MFC.
Honestly this experiment requires piles of agar plate. Like me, I got three sample extract and need to do triplicate for each of the sample. Sometimes for one extract, I need to uses 12 agar plates for one bacteria. Our group works on four types of bacteria and two fungi so just imagine how many plates are required...huhu last time we prepared 4 liters of SDA (1 liter can make up into 40-50 agar plates) and unfortunately it was overcooked so need to prepare it all over again...huhu I was totally devastated at that time because that incident happened three times in a row. To those whom doing antimicrobial, clean your apparatus well and distilled it. Check the autoclave's water level before autoclaving anything.
Okay I think that's all for antimicrobial part and insyaAllah I will post about antioxidant tests on the next entry.