Apr 15, 2025
Microplate characterisation
- 1CNRS@CREATE
Protocol Citation: sheena chan 2025. Microplate characterisation. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlkd47wg5r/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: April 15, 2025
Last Modified: April 15, 2025
Protocol Integer ID: 126701
Abstract
To characterise OD600 and GFP levels using a microplate reader.
Preparing pre-culture
Preparing pre-culture
Inoculate PGAL-GFP glycerol stock into a 50 mL falcon tube containing 5 mL Yeast Nitrogen Base (YNB) media containing 2% glucose (without leucine).
Incubate the sample overnight at 30 °C, with shaking.
Microplate characterisation workflow
Microplate characterisation workflow
After 22-24 h, measure the OD600 of the overnight pre-culture in a cuvette using Nanodrop OneC UV-Vis Spectrophotometer.
Refresh the overnight pre-culture with fresh YNB media containing 2% glucose (without leucine) to prepare four eppendorf tubes of 1 mL sample with final OD600 of 1.6.
Spin down the eppendorf tubes at 6,200 g for 1 min.
In each tube, resuspend the cell pellet with 1 mL of YNB media (without leucine) containing different galactose inducer concentrations:
A) 0.1% glucose (no galactose)
B) 0.1% glucose + 10 mM galactose
C) 0.1% glucose + 50 mM galactose
D) 0.1% glucose + 80 mM galactose
Plate 300 uL of the sample as triplicates in a clear 96-well plate and include well for media blank.
Load the plate into microplate reader (Cytation 5) to measure the OD600 and GFP level every 30 mins for 24 h.