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In addition to these quality filtering steps, a denoising step of the sequences was performed [ 17] with the "denoise_wrapper -script in qiime.
The lack of most quality filtering steps virtually prohibits the use of these other programs to perform GSPSD using the same initial error-prone data as used by POTION, with the chance of increasing the rate of false detection of positive selection to prohibitive rates [ 19].
After removing control features, four quality filtering steps were carried out sequentially using a range of quality control metrics produced by the Agilent Feature Extraction software to remove features that were saturated, non uniform, population outliers and spots non-significantly different from background.
After quality filtering steps and removal of outliers such as adaptor sequences and repeats.
After standard quality filtering steps, over 700 and 360 million 100 bp paired-end reads were obtained for X. couchianus and X. hellerii, respectively.
Additional sequencing was performed with a mixed cDNA pool on a Roche 454 FLX instrument, obtaining 274,607 high quality reads after trimming and quality filtering steps.
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At this point, another quality filtering step can be undertaken.
Both sequencing adapter and cDNA synthesis primers were removed in an additional quality filtering step.
For the reads that passed the quality filtering step, mapping was done using NOVOALIGN [ 59].
After the sequencing quality filtering step, a total of 424,743,516 of 100-base-pair Illumina reads were obtained for both samples (Table 1).
A total of 195, 088 HQ reads with an average sequence length of 356 bp were obtained from 202, 126 raw reads after the initial quality filtering step [NCBI Short Read Archive, accession SRA039388].
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