Oct 05, 2022
Determining biofilm growth amount (dry weight)
- 1XJTLU
Protocol Citation: Weizhuo.Chen, An.Huang 2022. Determining biofilm growth amount (dry weight). protocols.io https://dx.doi.org/10.17504/protocols.io.5jyl8jny9g2w/v1
License: This is an open access protocol 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
Protocol status: Working
We use this protocol and it's working
Created: September 19, 2022
Last Modified: October 05, 2022
Protocol Integer ID: 70204
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Abstract
This protocol will help determine the amount of biofilm grown on carriers.
IPTG induction
IPTG induction
Escherichia coli grown overnight was diluted by LB to OD600=0.4-0.6.
IPTG was added to cell culture to 1mM IPTG finally, and incubated 3h at 171 rpm, 37℃ in orbital shaking incubator.
Sample preparation
Sample preparation
For each bottle:
(1) The total mass of thirty Moving Bed Biofilm Reactor (MBBR) carriers were measured in advance by balance. Record all weights.
(2) Cells culture, thirty MBBR carriers and silver nitrate (to 6μM) solution were added into each bottle.
(3) Put all bottles into a 37℃ incubator.
Note
Prepare 3 flasks for each day and each sample (to minimize accidental error), so you may have, for example, 3 flasks for experiment group and 3 flasks for control group and you want to test for 5 days. This will eventually cost you 30 flasks in total.
Hot dry weight measurement
Hot dry weight measurement
For each day,
(1) MBBR carriers were dried by oven at 60℃.
(2) Paddings were cooled to room temperature.
(3) Total mass of thirty MBBR paddings were measured.
Repeat steps above until weight did not change. Record the weight.
The variation of MBBR carrier mass could be calculated, thus indicating the accumulated biomass (biofilm) on the carriers.