The total RNA of samples was isolated by RNAiso Plus reagent (Takara) according to the manufacturer's protocol. ReverTra Ace qPCR RT Kit (TaKaRa) was used to obtain cDNA. Quantitative reverse transcription polymerase chain reaction (qRT‐PCR) was performed using ABI 7500 StepOnePlus system (Applied Biosystems). β‐actin were used as internal controls and each reaction was performed in triplicate. All primers are shown in Supplemental file 1.
Abi 7500 steponeplus system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ABI 7500 StepOnePlus system is a real-time PCR (polymerase chain reaction) instrument designed for precise and efficient nucleic acid amplification and detection. It features a compact design, a touchscreen interface, and advanced software for data analysis and result interpretation. The system supports a variety of sample types and can be used for various applications, including gene expression analysis, pathogen detection, and genetic research.
Automatically generated - may contain errors
3 protocols using abi 7500 steponeplus system
1
RNA Isolation and qRT-PCR Analysis
2
Quantification of circRNA and mRNA Levels
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Total RNA was isolated by RNAiso Plus reagent (Takara, Otsu, Shiga, Japan) following the manufacturer’s protocol. One microgram of total RNA was reverse transcribed to cDNA using the PrimeScript RT Reagent Kit (Takara, Otsu, Shiga, Japan). The levels of circRNA and mRNA expression were measured by RT-PCR using SYBR Premix Ex Taq II (Takara) with the ABI 7500 StepOnePlus system (Applied Biosystems, CA, USA). β-actin was used as an internal control and each reaction was performed in triplicate. The primers are listed in Additional file 2 : Table S2.
3
Total RNA Extraction and qRT-PCR Analysis
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Total RNA was isolated from cells and tissues via RNAiso Plus reagent (Takara). The PrimeScript RT Reagent Kit (Takara) was used to reverse transcribe RNA to cDNA. The cDNAs were measured by quantitative reverse transcription–PCR using SYBR Premix Ex Taq II (Takara) with the ABI 7500 StepOnePlus system (Applied Biosystems). The primer sequences are provided in Supplemental Table 2 .
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