Trichoderma reesei conidia were used to inoculate 50 ml YPGel pre-cultivations. Cultivations were carried out at +28°C (200 rpm) for 24 h. Mycelium was centrifuged, supernatant removed and pellet resuspended in water. The suspensions were used to inoculate four ml SCD cultivations (three replicates each) in a 24-well plate to an initial density of 0.5 (OD600). Cells were cultivated 18 h at + 28°C (800 rpm) followed by centrifugation and resuspension in 600 μl of water. A total of 200 μl of the suspension was transferred to Black Cliniplate (Thermo Fisher Scientific) and mCherry fluorescence was measured with Varioskan in the same way as described above in the section ‘Fluorescence measurement of yeast species with fluorometer’. OD600 was measured by preparing 10× dilutions from the cell suspension. OD600 values were used for normalizing the fluorescence results to cell density.
Black cliniplate
Manufactured by Thermo Fisher Scientific
The Black Cliniplate is a laboratory equipment designed for various clinical applications. It serves as a multi-well plate for conducting assays, tests, and experiments in a controlled environment. The core function of the Black Cliniplate is to provide a standardized platform for sample preparation, incubation, and analysis.
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Lab products found in correlation
4 protocols using black cliniplate
1
Trichoderma reesei Conidia Cultivation
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Trichoderma reesei conidia were used to inoculate 50 ml YPGel pre-cultivations. Cultivations were carried out at +28°C (200 rpm) for 24 h. Mycelium was centrifuged, supernatant removed and pellet resuspended in water. The suspensions were used to inoculate four ml SCD cultivations (three replicates each) in a 24-well plate to an initial density of 0.5 (OD600). Cells were cultivated 18 h at + 28°C (800 rpm) followed by centrifugation and resuspension in 600 μl of water. A total of 200 μl of the suspension was transferred to Black Cliniplate (Thermo Fisher Scientific) and mCherry fluorescence was measured with Varioskan in the same way as described above in the section ‘Fluorescence measurement of yeast species with fluorometer’. OD600 was measured by preparing 10× dilutions from the cell suspension. OD600 values were used for normalizing the fluorescence results to cell density.
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Trichoderma reesei conidia were used to inoculate 50 ml YPGel pre-cultivations. Cultivations were carried out at +28°C (200 rpm) for 24 h. Mycelium was centrifuged, supernatant removed and pellet resuspended in water. The suspensions were used to inoculate four ml SCD cultivations (three replicates each) in a 24-well plate to an initial density of 0.5 (OD600). Cells were cultivated 18 h at + 28°C (800 rpm) followed by centrifugation and resuspension in 600 μl of water. A total of 200 μl of the suspension was transferred to Black Cliniplate (Thermo Fisher Scientific) and mCherry fluorescence was measured with Varioskan in the same way as described above in the section ‘Fluorescence measurement of yeast species with fluorometer’. OD600 was measured by preparing 10× dilutions from the cell suspension. OD600 values were used for normalizing the fluorescence results to cell density.
2
Fluorescent Yeast Strains Cultivation and Analysis
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Yeast cells (S. cerevisiae, Y. lipolytica, P. kudriavzevii, P. pastoris and C. apicola) were cultivated at +30°C on YPD plates for 24 h before starting the experiment, Z. lentus was cultivated at +24°C on YPD plates for 48 h. A total of 4 ml of SCD medium in 24-well plate was inoculated to initial optical density of 0.2 (OD600). Three parallel replicates were cultivated for each strain. Cells were cultivated for 18 h (+30°C/+24°C in case of Z. lentus, 800 rpm) followed by centrifugation and resuspension in 200 μl of water. A total of 200 μl of the suspension was transferred to Black Cliniplate (Thermo Fisher Scientific) and mCherry fluorescence was measured with Varioskan (Thermo Electron Corporation). The excitation and the emission wavelengths were 587/610 nm, measurement time was 500 ms using a 12 nm excitation bandwidth. Cell density (OD600) measurement was done for normalizing mCherry fluorescence measurement results. For this, cells were diluted 100× in water, and then OD600 was measured with Varioskan (photometric measurement mode, wavelength = 600 nm, bandwidth = 5 nm, measurement time = 100 ms) using a transparent microtiter plate (Nunc 96F, Thermo Fisher Scientific). Normalization was done by dividing the mCherry measurement value with OD600 result.
3
Fluorescent Protein Quantification
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Cultivation
samples were collected by centrifugation and resuspended with 200
μL of water. A 200 μL portion of the suspension was transferred
to Black Cliniplate (Thermo Fisher Scientific),, and fluorescence
was measured with Varioskan (Thermo Electron Corporation). The excitation/emission
wavelengths for measurement of BFP (blue fluorescent protein) were
399/456 nm, for Venus (yellow fluorescent protein) 510/530 nm, for
mCherry (red fluorescent protein) 587/610 nm, and for KO2 (orange
fluorescent protein) 550/570 nm, respectively. A 5 nm bandwidth and
500 ms measurement time were used in each measurement. Cell density
(OD600) measurement was done for normalizing fluorescence
measurement results. After fluorescence measurement, cells were diluted
100× into water, and then OD600 was measured with
Varioskan (photometric measurement mode, wavelength = 600 nm, bandwidth
= 5 nm, measurement time = 100 ms) using transparent microtiter plate
(Nunc 96F, Thermo Fisher Scientific). The arbitrary units (AU) reported
in figures were obtained by dividing the fluorescence measurement
value by the OD600 value.
samples were collected by centrifugation and resuspended with 200
μL of water. A 200 μL portion of the suspension was transferred
to Black Cliniplate (Thermo Fisher Scientific),, and fluorescence
was measured with Varioskan (Thermo Electron Corporation). The excitation/emission
wavelengths for measurement of BFP (blue fluorescent protein) were
399/456 nm, for Venus (yellow fluorescent protein) 510/530 nm, for
mCherry (red fluorescent protein) 587/610 nm, and for KO2 (orange
fluorescent protein) 550/570 nm, respectively. A 5 nm bandwidth and
500 ms measurement time were used in each measurement. Cell density
(OD600) measurement was done for normalizing fluorescence
measurement results. After fluorescence measurement, cells were diluted
100× into water, and then OD600 was measured with
Varioskan (photometric measurement mode, wavelength = 600 nm, bandwidth
= 5 nm, measurement time = 100 ms) using transparent microtiter plate
(Nunc 96F, Thermo Fisher Scientific). The arbitrary units (AU) reported
in figures were obtained by dividing the fluorescence measurement
value by the OD600 value.
4
Doxycycline-Induced Gene Expression Analysis
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Cells were cultivated
in duplicate on SCD-LEU-URA with 0, 1, 2, or 5 μg/mL doxycycline
for 20 h (30 °C, 800 rpm). Cells were centrifuged, resuspended
in 200 μL of DDIW, and transferred to Black Cliniplate (Thermo
Fisher Scientific). Venus (yellow fluorescent protein) fluorescence
was measured with Varioskan (Thermo Electron Corporation) using excitation
and emission wavelengths of 510/530 nm (measurement time = 100 ms).
A 100x dilution of the cell suspension was made for OD600 measurement with Varioskan (photometric measurement mode, wavelength
= 600 nm, bandwidth = 5 nm, measurement time = 100 ms) using a transparent
microtiter plate (Nunc 96F, Thermo Fisher Scientific), to normalize
fluorescence measurement originating from different cell densities.
The arbitrary units (AUs) reported in figures were obtained by dividing
the fluorescence measurement value by the OD600 value.
To monitor the effect of DOX on growth, the same cells were used
to inoculate 20 mL of SCD-URA-LEU medium, with 0, 1, 2, or 5 μg/mL
DOX. Cells were incubated at 30 °C at 200 rpm for around 17 h
with OD600 measurements taken regularly.
in duplicate on SCD-LEU-URA with 0, 1, 2, or 5 μg/mL doxycycline
for 20 h (30 °C, 800 rpm). Cells were centrifuged, resuspended
in 200 μL of DDIW, and transferred to Black Cliniplate (Thermo
Fisher Scientific). Venus (yellow fluorescent protein) fluorescence
was measured with Varioskan (Thermo Electron Corporation) using excitation
and emission wavelengths of 510/530 nm (measurement time = 100 ms).
A 100x dilution of the cell suspension was made for OD600 measurement with Varioskan (photometric measurement mode, wavelength
= 600 nm, bandwidth = 5 nm, measurement time = 100 ms) using a transparent
microtiter plate (Nunc 96F, Thermo Fisher Scientific), to normalize
fluorescence measurement originating from different cell densities.
The arbitrary units (AUs) reported in figures were obtained by dividing
the fluorescence measurement value by the OD600 value.
To monitor the effect of DOX on growth, the same cells were used
to inoculate 20 mL of SCD-URA-LEU medium, with 0, 1, 2, or 5 μg/mL
DOX. Cells were incubated at 30 °C at 200 rpm for around 17 h
with OD600 measurements taken regularly.
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