Total RNA was isolated using RNeasy kits (Qiagen, Hilden, Germany) primed with random hexamer oligonucleotides and was reverse transcribed using a PrimeScriptTM RT reagent kit (Takara, Japan). Real-time PCR was performed using an ABI 7300 Prism SDS real-time PCR detection system (Applied Biosystems, Foster City, CA, USA) with a SYBR® Premix Ex Tag kit (Takara Biotechnology, Shiga, Japan) and a standard temperature protocol. The results obtained using a cycle threshold are expressed as relative quantities and were calculated using the 2−ΔΔCT method (expressed as the relative fold ratio). All data were normalized to human GAPDH data, and three separate experiments were performed. Supplementary Table S2 lists the primers used during quantitative real-time PCR.
Abi 7300 prism sds real time pcr detection system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ABI 7300 Prism SDS real-time PCR detection system is a laboratory instrument used for real-time polymerase chain reaction (PCR) analysis. The system is designed to detect and quantify nucleic acid sequences in real-time during the amplification process.
Automatically generated - may contain errors
2 protocols using abi 7300 prism sds real time pcr detection system
1
Real-Time PCR RNA Quantification Protocol
2
Quantitative Real-Time PCR Protocol
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Total RNA was isolated from samples using QIAzol® RNA Lysis reagent (QIAGEN, Hilden, Germany), after which cDNAs were synthesized using a PrimeScript™ RT reagent Kit for RT-PCR (Takara Bio Inc., Otsu, Japan) according to the manufacturer’s instructions. Quantitative PCR was performed using an ABI 7300 Prism SDS real-time PCR detection system (Applied Biosystems, Foster City, CA, USA) with a SYBR® Premix Ex Tag kit (Takara Bio Inc., Otsu, Japan) and a standard temperature protocol. The results obtained using CT (cycle threshold) were expressed as relative quantities and were calculated using the 2-ΔΔCT method (expressed as relative fold ratio). Hypoxanthine phosphoribosyltransferase 1 (HPRT1) was used as a control gene for normalization, and three separate experiments were performed. Detailed primer information is shown in Figure 2 .
Primers used for the quantitative real-time PCR ![]()
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