Nov 20, 2020

Public workspaceCalculating Colony-forming Units

DOI
https://dx.doi.org/10.17504/protocols.io.bnuvmew6
Calculating Colony-forming Units
  • Jiri Hulcr1,
  • You Li1
  • 1University of Florida
  • Protocols Bark Beetle Mycobiome
Bark Beetle Mycobiome Research Coordination Network Bark
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DOI: https://dx.doi.org/10.17504/protocols.io.bnuvmew6
Document Citation: Jiri Hulcr, You Li 2020. Calculating Colony-forming Units. protocols.io https://dx.doi.org/10.17504/protocols.io.bnuvmew6
License: This is an open access document distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Created: October 23, 2020
Last Modified: November 20, 2020
Document Integer ID: 43637
Keywords: bark beetle mycobiome, part of the bark beetle mycobiome, fungus symbiosis, number of colony, more information on the bbm, calculating colony, widespread insect, symbiosis, fungus, bbm, colony
Abstract
This protocol describes how to estimate the number of colony-forming units.

This protocol is part of the Bark Beetle Mycobiome (BBM) Research Coordination Network. For more information on the BBM international network: Hulcr J, Barnes I, De Beer ZW, Duong TA, Gazis R, Johnson AJ, Jusino MA, Kasson MT, Li Y, Lynch S, Mayers C, Musvuugwa T, Roets F, Seltmann KC, Six D, Vanderpool D, & Villari C. 2020. Bark beetle mycobiome: collaboratively defined research priorities on a widespread insect-fungus symbiosis. Symbiosis 81: 101–113 https://doi.org/10.1007/s13199-020-00686-9.
Troubleshooting
To estimate the initial number of colony-forming units in a mycangium, use the technique of serial dilution when plating. After colonies grow, multiply the number of colonies on the plate by the inverse of the initial dilution factor.

  1. Prepare two tubes per each sample, label them “0.1” and “0.01”, fill each with 500 ul of water or PBS.
  2. Suspend mycangium in the tube “0.1” and vortex.
  3. Plate 50 ul of the suspension on media. Record the plate as “0.1 dilution” in [PLATES].[note] in the Isolations database.
  4. Transfer 50 ul of the initial suspension to the second tube (“0.01”) and vortex.
  5. Plate 50 ul of the second suspension on second media plate, and record that plate as “0.01 dilution”.
  6. Plate 5 ul of the second suspension on a third plate, and record it as “0.001 dilution”.