qPCR data for reference gene selection in tomato leaf tissue infected with Tomato curly stunt virus

Published: 23 January 2020| Version 1 | DOI: 10.17632/ccb5mkpkz5.1
Contributors:
Farhahna Allie,
Imanu Mwaba,
Mamokete Bokhale

Description

qPCR data for Actin7 (ACT), Β-6 Tubulin (TUB), Ubiquitin 3 (UBI), Clathrin adaptor complexes medium subunit (CAC), Phytoene desaturase (PDS), Expressed protein (EXP), Glyceraldehyde-3-phosphate dehydrogenase(GAPDH), Adenine phosphoribosyl transferase-like protein (APT1), TAP42-interacting protein (TIP41) and Elongation factor 1-alpha (EF1α) , measure in tomato leaf tissue infected with Tomato curly stunt virus.

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Tomato gene sequences for Actin7 (ACT), Β-6 Tubulin (TUB), Ubiquitin 3 (UBI), Clathrin adaptor complexes medium subunit (CAC), Phytoene desaturase (PDS), Expressed protein (EXP), Glyceraldehyde-3-phosphate dehydrogenase(GAPDH), Adenine phosphoribosyl transferase-like protein (APT1), TAP42-interacting protein (TIP41) and Elongation factor 1-alpha (EF1α) were obtained from the Sol Genomics network (https://solgenomics.net/). FASTA sequence for each gene was used for qPCR prime design using the intergrated DNA technologies PrimerQuest tool. RNA was extracted from resistant and Susceptible leaf tissue infected with Tomato curly stunt virus RNA miniprep kit (Zymo Research, USA) as per manufacturers instruction. RNA was reverse transcribed to cDNA using Maxima H Minus First Strand cDNA Synthesis kit RT-qPCR (Thermo scientific, USA). ). Primer efficiencies were obtained for each candidate genes using dilutions of cDNA and a standard curve was generated from five 10-fold dilution series on The CFX connect™ Real-Time System (Biorad, USA). Melt curve analysis was conducted for each candidate gene using the Bio-Rad CFX manager 3.1