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The variation in the quantity of identified peaks indicates that default stringency levels are tuned differently among programs.
The default stringency settings were used: maximum number of allowed UN-base paired bases = 0 in seed/nucleus of 7 nucleotides, and minimum number of paired-up bases in heteroduplex = 14; maximum folding energy for heteroduplex (Kcal/mol = −25).
The default stringency setting of the P/A call criteria used above was meant to control the false present calls rate at 0.05, i.e., an expected false present call rate of 5%.
This was the result of default stringency levels causing parts of some sequences with low quality bases to be discarded.
In addition, Harr or dot plots were constructed using Pipmaker [ 74], under default stringency settings, to compare potentially homologous regions of the Welwitschia genome with gene sequences from the plastomes of Podocarpus macrophyllus and Ginkgo biloba (Raubeson et al, unpublished).
The sequence data generated from each region, corresponding to each of the 16 groups, was assembled separately using the GS Amplicon Variant Analyzer software (Roche, 454 Life Sciences) with default stringency settings.
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Single-nucleotide polymorphisms (SNPs) were then called using the diBayes algorithm with conservative default call stringency [ 8].
"Functional Annotation Clustering" with default classification stringency is applied to derive all the related functions associating with their enrichment scores and P-values.
Burrows-Wheeler Aligner (BWA) version 0.7.5a-r405 [ 97] was used to align the unprocessed Illumina reads to the EST library using the default alignment stringency.
The annotations have been selected based on medium stringency (default) setting, with > 2-fold enrichment and Benjamini-Hochberg p-value < 0.05.
The DAVID annotation clustering module [ 25, 26] set at medium stringency (default level) was used to classify differentially expressed G. nigrifrons transcripts into functional groups.
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