RNA was extracted from cultured cells using the Zymo Quick RNA Miniprep Kit following the manufacturer’s protocol. Reverse transcription (RT) reactions were performed as described (Somarelli et al. 2016) using the ABI High Capacity cDNA Reverse Transcription Kit (ThermoFisher). RT reactions were incubated following the manufacturer’s protocol in a SimpliAmp thermocycler (Life Technologies). RT reactions were diluted 1:5 in nuclease-free H2O, and RT-qPCR was performed as described (Somarelli et al. 2016) in a Vii7 real time-PCR detection system (Applied Biosystems).
Vii7 real time pcr detection system
Manufactured by Thermo Fisher Scientific
The Vii7 real-time PCR detection system is a laboratory instrument designed for the amplification and detection of DNA sequences. It is capable of performing real-time quantitative PCR (qPCR) analyses. The system includes optical components, temperature control, and software for data acquisition and analysis.
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Lab products found in correlation
3 protocols using vii7 real time pcr detection system
1
RNA Extraction, RT, and qPCR Protocol
2
Quantitative RT-PCR Protocol for Gene Expression
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For qPCR, total RNA was reverse transcribed using the High-Capacity cDNA Reverse Transcription Kit (Life Technologies). Aliquots of 5-fold diluted reverse transcription reactions were subjected to quantitative (q)PCR with KAPA SYBR FAST master mix using the Vii7 real time-PCR detection system (Applied Biosystems). GAPDH mRNA levels were measured for normalization, and the data are presented as “Relative Expression”.
3
Quantitative Real-Time PCR Protocol
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Total RNA was isolated using the Quick-RNA Miniprep kit (Zymo Research). Total RNA was reverse transcribed using the High-Capacity cDNA Reverse Transcription Kit (Life Technologies). Aliquots of 5-fold diluted reverse transcription reactions were subjected to quantitative (q)PCR with KAPA SYBR FAST master mix using the Vii7 real time-PCR detection system (Applied Biosystems). GAPDH mRNA levels were measured for normalization, and the data are presented as “Relative Expression”. A complete list of primer sequences is provided in the Supplementary Text.
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