MicroStation System, MicroLog Version 4.2 - DTU Systems Biology ...

Preparing Samples
Caution!
Pipet the
inoculum into a
MicroPlate
within
20 minutes!
Use sterile
technique.
2. Do NOT add a source of moisture to this container. Mycelial growth
will overwhelm the agar plate and cause crossover contamination of
other cultures.
3. Incubate the 2% ME agar at 26° C for 5-10 days to induce conidia
formation (incubate longer if required). Incubate yeasts for 1-2 days.
Inoculating MicroPlates
Inoculate the suspension into one of the following Biolog MicroPlates:
• GN2: For all gram-negative aerobic bacteria
Caution!
Accurately
pipet
recommended
volumes.
• GP2: For all gram-positive aerobic bacteria
• AN: For all anaerobic bacteria
• YT: For all yeasts visceral
• FF: For all filamentous fungi and select yeast strains
Pipet the inoculum into a Biolog MicroPlate within 20 minutes. Waiting
any longer may cause inaccurate identification. Anaerobic bacteria are
especially sensitive to delays. If you are running a batch of MicroPlates,
set them up (from preparing the inoculum to pipetting into the MicroPlate)
so you will not exceed the 20 minute deadline.
Inoculating protocol
1. Label the MicroPlate with the organism name/number and plate type
(e.g., GN, GP). Label the side of the MicroPlate itself, not the lid.
2. Using aseptic technique, pour the cell suspension into a multichannel
pipette reservoir.
3. Firmly attach eight sterile tips to the pipettor.
4. Fill the tips with the suspension. Check to see that all tips fill equally.
5. Prime the tips by dispensing once back into the reservoir. The
electronic pipettor will do this automatically.
6. Fill all MicroPlate wells by placing the tips at an angle inside the top
of the wells . Take care not to splash from one well to another. Avoid
contamination. Avoid touching the bottom of the wells, which could
transfer carbon sources.
• Gram negatives and gram positives � 150 µl per well
• Anaerobes, yeasts and filamentous fungi � 100 µl per well
Section 4 � Page 14 MicroStation System/MicroLog User Guide Apr 07