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Applied biosystems 7500 thermal cycler

Manufactured by Thermo Fisher Scientific

The Applied Biosystems 7500 thermal cycler is a piece of laboratory equipment designed for nucleic acid amplification. It is capable of performing polymerase chain reaction (PCR) and other thermal cycling-based techniques. The 7500 thermal cycler provides precise temperature control and consistent performance for a variety of sample types and applications.

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2 protocols using applied biosystems 7500 thermal cycler

1

Thermal Shift Assay of BOK and BAK Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
TSAs were performed by using the Protein Thermal Shift Starter Kit
(ThermoFisher), according to the manufacturer’s protocol, using 0.25
mg/mL purified BOK and BAK. Briefly, for each protein sample, a 90 μL
MasterMix was prepared by using the buffer and the fluorescent dye ROX,
vortexed at low speed, and 4 aliquots of 20 μL each were dispensed in
4 individual wells of Microamp Fast Optical 96-well reaction plates
(ThermoFisher) for analysis in an Applied Biosystems 7500 thermal cycler
(ThermoFisher). The temperature gradient was 25°C–99°C
at a rate of 1°C/min. Data were analyzed with Protein Thermal Shift
Software v1.3 (ThermoFisher) to determine the melting temperature, using the
first derivative and Boltzmann equation non-linear regression. Dye-only
controls were included in each plate. Experiments were performed at least 2
times in quadruplicates for each protein sample at the 2 protein
concentrations. More recently, TSA assays were performed in 20 mM NaOAc pH
5.0 or 20 mM HEPES pH 6.8 and 150 mM NaCl, and 384-well plates in an Applied
Biosystems 7900HT Fast Real-Time PCR System (ThermoFisher). Final data
analysis and presentation were done in Microsoft Excel and GraphPad Prism v7
(GraphPad Software).
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2

Thermal Shift Assay of BOK and BAK Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
TSAs were performed by using the Protein Thermal Shift Starter Kit
(ThermoFisher), according to the manufacturer’s protocol, using 0.25
mg/mL purified BOK and BAK. Briefly, for each protein sample, a 90 μL
MasterMix was prepared by using the buffer and the fluorescent dye ROX,
vortexed at low speed, and 4 aliquots of 20 μL each were dispensed in
4 individual wells of Microamp Fast Optical 96-well reaction plates
(ThermoFisher) for analysis in an Applied Biosystems 7500 thermal cycler
(ThermoFisher). The temperature gradient was 25°C–99°C
at a rate of 1°C/min. Data were analyzed with Protein Thermal Shift
Software v1.3 (ThermoFisher) to determine the melting temperature, using the
first derivative and Boltzmann equation non-linear regression. Dye-only
controls were included in each plate. Experiments were performed at least 2
times in quadruplicates for each protein sample at the 2 protein
concentrations. More recently, TSA assays were performed in 20 mM NaOAc pH
5.0 or 20 mM HEPES pH 6.8 and 150 mM NaCl, and 384-well plates in an Applied
Biosystems 7900HT Fast Real-Time PCR System (ThermoFisher). Final data
analysis and presentation were done in Microsoft Excel and GraphPad Prism v7
(GraphPad Software).
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