A549 adenocarcinoma cells were cultured in RPMI1640 media containing 10% fetal bovine serum, 2mM glutamine, 50 units/ml penicillin, and 50 μg/ml streptomycin (all from Life Technology, Inc.), at 37°C, 5% CO2, and 100% humidity. For screens, cells were grown in 10 cm culture plates overnight, detached by trypsin, counted by TC10 automated cell counter (Bio-Rad Laboratories, Inc.) and seeded onto 384-well plate at a density of 3000 cells/well by Matrix electronic multichannel pipette (Matrix Technologies Co.). After 24 hrs at 37°C, drugs were added by Beckman Coulter BioMek FX liquid handlers (Beckman Coulter, Inc.), and the plate was covered by Breath-Easy sealing membrane (Sigma-Adrich, Inc.) and incubated at 37°C for two days.
Breath easy sealing membrane
Manufactured by Merck Group
Sourced in Switzerland
The Breath-Easy sealing membrane is a lab equipment product from Merck Group. It is a semipermeable membrane that allows the passage of gases while preventing the transfer of liquids and microorganisms. The core function of this product is to provide a controlled and secure environment for various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Biomek fx, by Beckman Coulter (2 mentions)
Tc10 automated cell counter, by Bio-Rad (2 mentions)
Infinite m200 pro plate reader, by Tecan (1 mentions)
Flat bottom 96 well microtiter plate, by Greiner (1 mentions)
4 protocols using breath easy sealing membrane
1
High-Throughput Drug Screening in A549 Cells
2
Hygromycin Sensitivity of Ai and Se
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Hygromycin sensitivity of Ai and Se was measured in liquid cocultures. Duplicate co-cultures were initiated by mixing purified Se with Ai at an OD600 of 0.2 in TSY and at an approximate cellular ratio of 1:2 (Se:Ai). The co-cultures were divided into multiple aliquots in 96-well culture plates and hygromycin was added to the final concentrations shown in Figure S1 . The plates were covered with Breath-Easy sealing membrane (Sigma Z380059) and incubated without lids at 37°C with 5% CO2. At 1 day and 3 days, the cells within individual wells were pelleted (>15 min at >15,000 rcf) and stored at −20°C for later genomic DNA isolation and qPCR-based quantification of Se and Ai (see below).
3
High-Throughput Drug Screening in A549 Cells
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A549 adenocarcinoma cells were cultured in RPMI1640 media containing 10% fetal bovine serum, 2mM glutamine, 50 units/ml penicillin, and 50 μg/ml streptomycin (all from Life Technology, Inc.), at 37°C, 5% CO2, and 100% humidity. For screens, cells were grown in 10 cm culture plates overnight, detached by trypsin, counted by TC10 automated cell counter (Bio-Rad Laboratories, Inc.) and seeded onto 384-well plate at a density of 3000 cells/well by Matrix electronic multichannel pipette (Matrix Technologies Co.). After 24 hrs at 37°C, drugs were added by Beckman Coulter BioMek FX liquid handlers (Beckman Coulter, Inc.), and the plate was covered by Breath-Easy sealing membrane (Sigma-Adrich, Inc.) and incubated at 37°C for two days.
4
Growth Kinetics of S. bombicola Strains
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Specific growth rates for S. bombicola strains were determined by incubating 200 μL of cell culture in Lang medium in a flat bottom, 96-well microtiter plate (Greiner Bio-One) at 30 °C in a Tecan Infinite® m200 Pro plate reader (Tecan, Mannedorf, Switzerland) with an initial OD600 of 0.05, sealed off with a Breath-Easy® sealing membrane (Sigma-Aldrich). The OD600 was measured every 15 min for 40 h, while shaking in between measurements at 200 rpm (orbital mode, amplitude 2 mm). The maximum specific growth rate (μ) was calculated by fitting the Richards model in with the ‘curve_fit ‘function in the ‘SciPy’ module in Python 3.7 [65 (link)].
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