Animals were synchronized and total RNA extraction was done following the protocol described above. qRT-PCR was conducted using the Applied Biosystems One-Step Real-time PCR protocol using SYBR Green fluorescence (Applied Biosystems) on an Applied Biosystems 7900HT real-time PCR machine in 96-well-plate format. Twenty-five microliter reactions were analyzed as outlined by the manufacturer (Applied Biosystems). The relative fold changes of the transcripts were calculated using the comparative CT(2-ΔΔCT) method and normalized to pan-actin (act-1, −3, −4). The cycle thresholds of the amplification were determined using StepOnePlus™ Real-Time PCR System Software v2.3 (Applied Biosystems). All samples were run in triplicate. The primer sequences were available upon request and presented in Table S6 .
Steponeplus real time pcr system software v2
Manufactured by Thermo Fisher Scientific
The StepOnePlus Real-Time PCR System Software v2.3 is a software application designed to control the StepOnePlus Real-Time PCR System. It provides the interface and tools necessary to run and analyze real-time PCR experiments. The software includes features for experiment setup, data collection, and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Sybr green fluorescence, by Thermo Fisher Scientific (7 mentions)
One step real time pcr protocol, by Thermo Fisher Scientific (5 mentions)
Script reverse transcription supermix kit, by Bio-Rad (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Power sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions)
8 protocols using steponeplus real time pcr system software v2
1
Quantitative Real-Time PCR Analysis
2
Quantitative RT-PCR Analysis of Gene Expression
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Animals were synchronized and total RNA extraction was done following the protocol described above. qRT-PCR was conducted using the Applied Biosystems One-Step Real-time PCR protocol using SYBR Green fluorescence (Applied Biosystems) on an Applied Biosystems 7900HT real-time PCR machine in 96-well-plate format. Twenty-five-microliter reactions were analyzed as outlined by the manufacturer (Applied Biosystems). The relative fold-changes of the transcripts were calculated using the comparative CT(2−ΔΔCT) method and normalized to pan-actin (act-1, −3, −4). The cycle thresholds of the amplification were determined using StepOnePlus Real-Time PCR System Software v2.3 (Applied Biosystems). All samples were run in triplicate. The primer sequences were available upon request and presented in Table S8 .
3
Quantifying Gene Expression by qRT-PCR
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Animals were synchronized, and total RNA extraction was performed following the above-described protocol. qRT-PCR was conducted using the Applied Biosystems One-Step Realtime PCR protocol with SYBR Green fluorescence (Applied Biosystems) on an Applied Biosystems 7900HT real-time PCR machine in 96-well plate format. The reactions were analyzed as outlined by the manufacturer (Applied Biosystems). The relative fold changes of the transcripts were calculated using the comparative CT(2−ΔΔCT) method and normalized to pan-actin (act-1, -3, -4). The cycle thresholds of the amplification were determined using StepOnePlus Real-Time PCR System Software v2.3 (Applied Biosystems). All samples were run in triplicate. The primer sequences are presented in S10 Table .
4
Quantitative Gene Expression Analysis
5
Quantitative Real-Time PCR Analysis
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Animals were synchronized and total RNA extraction was done following the protocol described above. qRT-PCR was conducted using the Applied Biosystems One-Step Real-time PCR protocol using SYBR Green fluorescence (Applied Biosystems) on an Applied Biosystems 7900HT real-time PCR machine in 96-well-plate format. Twenty-five microliter reactions were analyzed as outlined by the manufacturer (Applied Biosystems). The relative fold changes of the transcripts were calculated using the comparative CT(2-ΔΔCT) method and normalized to pan-actin (act-1, -3, -4). The cycle thresholds of the amplification were determined using StepOnePlus™ Real-Time PCR System Software v2.3 (Applied Biosystems). All samples were run in triplicate. The primer sequences were available upon request and presented in Table S7 .
6
Quantitative RT-PCR Protocol for Transcriptome Analysis
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Animals were synchronized and total RNA extraction was done following the protocol described above. Quantitative reverse transcription-PCR (qRT-PCR) was conducted using the Applied Biosystems One-Step Real-time PCR protocol using SYBR Green fluorescence (Applied Biosystems) on an Applied Biosystems 7900HT real-time PCR machine in 96-well plate format. Twenty-five microliter reactions were analyzed as outlined by the manufacturer (Applied Biosystems). The relative fold changes of the transcripts were calculated using the comparative CT(2-ΔΔCT) method and normalized to pan-actin (act-1, –3, –4). The cycle thresholds of the amplification were determined using StepOnePlus Real-Time PCR System Software v2.3 (Applied Biosystems). All samples were run in triplicate. The primer sequences were available upon request and presented in Supplementary file 7 .
7
Quantitative RT-PCR Analysis of Gene Expression
8
Comparative qRT-PCR Analysis of Gene Expression
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Animals were synchronized and total RNA extraction was done following the protocol described above. qRT-PCR was conducted using the Applied Biosystems One-Step Real-time PCR protocol using SYBR Green fluorescence (Applied Biosystems) on an Applied Biosystems 7900HT real-time PCR machine in 96-well-plate format. Twenty-five-microliter reactions were analyzed as outlined by the manufacturer (Applied Biosystems). The relative fold-changes of the transcripts were calculated using the comparative CT(2 -ΔΔCT ) method and normalized to pan-actin (act-1, -3, -4). The cycle thresholds of the amplification were determined using StepOnePlus™ Real-Time PCR System Software v2.3 (Applied Biosystems). All samples were run in triplicate.
The primer sequences were available upon request and presented in Table S9.
The primer sequences were available upon request and presented in Table S9.
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