add qPCR assay and protocol

assays/qRT-PCR/protocols/VerificationOfGeneExpressionUsingqRT-PCR.md

+## Verification of gene expression using qRT-PCR
+
+To verify the overall consistency of our estimated TPM values in this metaanalysis, we performed qRT-PCR on twelve NBS-LRR-genes and three reference genes (de Vries et al., 2018). We evaluated the expression of these fifteen genes over six time points on the Moneymaker cultivar inoculated with *Phytophthora infestans (P. infestans)* isolate IPO-C. Three replicates were studied at each sampling time point and treatment type. Additional details of this experiment are reported in de Vries et al., 2018. The Bioproject L (PRJNA487149) from Fawke et al., 2019 is the most similar in design to our 2018 study, since that project sampled transcriptomes from leaves of the tomato cultivar ‘MicroTom’ inoculated with *P. infestans* isolate 88069. We evaluated the consistency between the average TPM of these 15 genes from Fawke et al. with our estimated Cq values at 72 hours post infection, the single overlapping timepoint between both data sets.
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