Total RNA was extracted by the TRIZOL method, and the first cDNA strand was generated using Toyobo cDNA First Strand Synthesis Kit (Toyobo, Shanghai, China). Then, the RT-qPCR was performed on the Roche LightCycler®480 Real-Time PCR instrument (Rotkreuz, Switzerland) with Toyobo Premix Kit (Toyobo, Shanghai, China). Three independent biological replicates and three technical replicates were adopted. The N. benthamiana ubiquitin-conjugating enzyme E2 (UBC) gene was used as the internal reference gene to calculate relative gene expression levels using the 2−ΔΔ C(t) method [49 (link)]. All primers are listed in the Supplementary Table S4 .
Lightcycler 480 real time pcr instrument
Manufactured by Toyobo
Sourced in Switzerland
The LightCycler®480 Real-Time PCR instrument is a laboratory equipment used for real-time polymerase chain reaction (PCR) analysis. It provides a platform for quantitative gene expression analysis, genotyping, and other real-time PCR applications.
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Lab products found in correlation
3 protocols using lightcycler 480 real time pcr instrument
1
Quantifying gene expression in N. benthamiana
2
Quantitative Gene Expression Analysis
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Total RNA was extracted by the TRIZOL method, and 1 μg total RNA was used for reverse transcription using Toyobo cDNA First Strand Synthesis Kit. Subsequently, RT-qPCR was performed on the Roche LightCycler® 480 Real-Time PCR instrument with Toyobo Premix Kit. Three independent biological replicates and three technical replicates were adopted. The T. aestivum cell division cycle (CDC) gene (accession number XM_020313450) was used as the internal reference gene, and the relative expression of the gene was calculated by the 2−ΔΔC(t) method [41 (link)]. All primers are listed in the Supplementary Table S4 .
3
Total RNA Extraction and RT-qPCR Analysis
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Total RNA was extracted by the TRIZOL method, and 1 μg total RNA was used for reverse transcription using the Toyobo cDNA First Strand Synthesis Kit. RT-qPCR was then performed on a Roche LightCycler®480 Real-Time PCR instrument with Toyobo Premix Kit. Three independent biological replicates with three technical replicates were performed. All primers are listed in the Additional file 6 : Table S5.
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