Total RNA was isolated using TRIzol reagent (BIOO Scientific Co., Austin, TX, USA) according to the manufacturer’s protocol. RNA was reverse‐transcribed using M‐MLV Reverse Transcriptase (Promega Inc., Madison, WI, USA), and SYBR green‐based quantitative real‐time PCR (Vazyme Biotech, Nanjing, China) was subsequently performed. Expression analysis was performed using specific primers, and primer sequences are shown in Table S1 .
Sybr green based quantitative real time pcr
Manufactured by Vazyme
Sourced in United States, China
SYBR green-based quantitative real-time PCR is a molecular biology technique used for the quantification of specific DNA sequences. It utilizes the SYBR green dye, which binds to double-stranded DNA, to detect and measure the amplification of target DNA in real-time during the polymerase chain reaction (PCR) process.
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2 protocols using sybr green based quantitative real time pcr
1
Total RNA Isolation and qRT-PCR Analysis
2
Investigating Immune Responses in Porcine Cell Lines
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Total RNA was extracted from 293T, WSL and PK15 cells using Simple P Total RNA Extraction Kit (Bioer Technology, China) according to the manufacturer's instructions. Samples were subjected to reverse transcription using HiScript II Q RT kit (Vazyme Biotech, China). The cDNA was used as a template for Semi-quantitative RT-PCR to investigate the mRNA levels of pig IL-1, IL-6, IL-8, IFN, TNF, D345L and Actin. The mRNA levels of IFN and GAPDH were quantitated by SYBR green-based quantitative real-time PCR (Vazyme Biotech, China) using a Life Technology instrument. The primers used here were listed in Table 1.
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