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Modules were grouped using a stringency threshold of 0.75.
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In other words, using the trend for nonspecific genes at lower stringency levels, a stringency threshold was chosen that was high enough to consider 90% of the genes highly expressed in either fetal liver or placenta to be specifically derived from that tissue and not be a statistical artefact.
With a stringency threshold of 0.75, a total of 10 modules were identified from petunia corollas using WGCNA (Additional file 3).
To overcome the relatively high stringency imposed by the maximal clique method, the Quasi Clique [ 7], Molecular Complex Detection (MCODE) [ 12], Spirin and Mirny [ 11] algorithms identify densely connected subgraphs rather than fully connected ones by either optimizing an objective density function or using a density threshold.
After hybridization the membranes were washed twice using a low stringency buffer (2× SSC, 0.1% SDS), and a high stringency buffer (0.1× SSC, 0.1% SDS), for 15 and 5 minutes at 48°C, respectively.
As with Tm-matched probe-sets, processing microarrays at multiple stringencies resulted in higher success rates than when using a common stringency (compare strategy 5 and 8 in Figure 4).
Amplification was performed in a thermal cycler (Appendorf) using a high stringency PCR protocol with high annealing temperature to ensure specificity of amplification.
The CB maps for each contig with 2 or more Q clones were recalculated using a higher stringency cutoff value.
SNPs were called using a simple thresholding.
The absence of NPM-related proteins in thirty-seven diverse plant species with sequenced genomes was confirmed using PLAZA 3.0 Dicot and PLAZA 3.0 monocot comparative genomics platforms (http://bioinformatics.psb.ugent.be/plaza/) [ 60] using BLASTp at a low stringency threshold (expect: 1).
using an arbitrary threshold.
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