Total RNA was isolated using a NucleoSpin® RNA kit (MACHEREY-NAGEL) following the user’s instruction. First-strand cDNAs were synthesized from RNase-free DNase I treated (Invitrogen, Waltham, MA, USA) total RNA to eliminate genomic DNA contamination using a High Capacity cDNA synthesis™ system (Applied Biosystems, Waltham, MA, USA). qRT-PCR was performed with ABI Prism 7900 HT Real-time PCR system (Applied Biosystems, Waltham, MA, USA) using Power SYBR® Green PCR Master Mix (Applied Biosystems, Waltham, MA, USA). Three replicates were performed for the analysis of each sample. Potato 18S-rRNA as an internal control and the relative expression levels of the target gene were determined. For relative quantification, the 2−ΔΔCT method between conditions in RT-qPCR was applied. The sequence of the primers is listed in Supplementary Table S6 .
High capacity cdna synthesis system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The High Capacity cDNA synthesis™ system is a laboratory instrument designed for efficient and reliable conversion of RNA into complementary DNA (cDNA). The system provides a streamlined workflow for the production of high-quality cDNA samples from a variety of RNA sources.
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2 protocols using high capacity cdna synthesis system
1
RNA Isolation and qRT-PCR Analysis
2
RNA Isolation and qRT-PCR Analysis
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Total RNA was isolated using a NucleoSpin® RNA kit (MACHEREY-NAGEL) following the user’s instruction. First-strand cDNAs were synthesized from RNase-free DNase I treated (Invitrogen, Waltham, MA, USA) total RNA to eliminate genomic DNA contamination using a High Capacity cDNA synthesis™ system (Applied Biosystems, Waltham, MA, USA). qRT-PCR was performed with ABI Prism 7900 HT Real-time PCR system (Applied Biosystems, Waltham, MA, USA) using Power SYBR® Green PCR Master Mix (Applied Biosystems, Waltham, MA, USA). Three replicates were performed for the analysis of each sample. Potato 18S-rRNA as an internal control and the relative expression levels of the target gene were determined. For relative quantification, the 2−ΔΔCT method between conditions in RT-qPCR was applied. The sequence of the primers is listed in Supplementary Table S6 .
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