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Therefore, another software program, RefFinder (Radonić et al. 2004), was used to integrate the currently available major computational programs (geNorm, NormFinder, BestKeeper, and the comparative ΔΔCt method) to compare and rank the tested candidate reference genes.
Sequences were selected from GenBank to represent the available major vertebrate groups (i.e., fish, amphibians, and mammals).
For this purpose, most of the publicly available major sequence assemblers – both for low-coverage long (Sanger) and high-coverage short (Illumina) reads technologies – are compared.
To reinforce this result, RefFinder (http://www.leonxie.com/referencegene.php), a web-based tool that integrates the currently available major computational programs (geNorm [ 33], Normfinder [ 32], BestKeeper [ 34] and the comparative ΔCt method [ 72]) was also used.
The mining of diverse loci and gene functions controlling polygenic resistance, combined and deployed together with available major resistance genes, is expected to be a key strategy for durably improving plant genetic resistance to pathogens.
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