The qRT-PCR was performed in an iQTM5 multicolor RT-PCR detection system (Bio-Rad), in a 10 μl reaction volume using the ssoFast EvaGreen Supermix (Bio-Rad). The sequences of primers used for the qRT-PCR for RNA-seq validation are listed in Supplementary Table S2 . The efficiency of the PCR was calculated using the program LinRegPCR (40 (link)). Normalized data were calculated by dividing the average of at least three replicates of each sample from the candidate and the reference gene, rnpB.
Iqtm5 multicolor rt pcr detection system
Manufactured by Bio-Rad
Sourced in United States
The IQ5 multicolor RT-PCR detection system is a real-time PCR instrument manufactured by Bio-Rad. It is designed to perform quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis with up to five fluorescent dyes simultaneously.
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Lab products found in correlation
2 protocols using iqtm5 multicolor rt pcr detection system
1
qRT-PCR for RNA-seq Validation
2
Quantitative Gene Expression Analysis
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RNA was extracted from cell pellets using Total RNA Extraction Kit (Solarbio Life Science, Beijing, China). cDNA was then synthesized from 1 µg of RNA using Solarbio's Maxima First Strand cDNA Synthesis Kit (Catalog #K1672) and amplified by qRT-PCR on iQTM5 Multicolor RT-PCR Detection System (Bio-Rad, Hercules, CA, USA) using PowerUp ™ SYBR ™ Green Master Mix (Applied Bioscience, Carlsbad, CA, USA) and Bio-Rad's PrimePCR ™ primer pairs for TrxR1 and GPX1. Relative quantification of gene expression was calculated using the 2 -ΔΔCT method with GAPDH serving as a housekeeping gene (Alfhili et al. 2021) (link).
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