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Color channels were carefully aligned using alignment parameter from control measurements with 0.5 µm diameter multi-spectral fluorescent beads (Life technologies).
Images for each fluorophore were acquired simultaneously with independent EMCCD cameras (iXON; Andor), and images were aligned post-capture using alignment parameters generated from images of 0.1-µm fluorescent microspheres (TetraSpeck; Invitrogen).
Using alignment database and Bowtie, SpliceSeq aligns reads from RNA-Seq data to a reference collection of splice variants [ 19, 20].
Using a visual browser such as Ensembl, these methylated reads (sequences) are aligned and compared to the human genome using alignment software such as MeQA (http://life.tongji.edu.cn/meqa/) (Huang et al., 2012).
The lack of a pig genome draft assembly still hampers the traditional method of identifying SNPs, in which DNA shotgun sequences of different individuals are aligned to a genomic region of interest using alignment algorithms [ 14].
Without using alignment thresholds, more than 99.9% of all sequences could be aligned back to the reference.
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However, only 4/8 of the sets were correctly classified using alignment-based cross-species conservation.
Surprisingly, the optimal recall rate using alignment-based conservation was only 25/70 (36%) at 10% DNA coverage, showing no improvement.
Maybe the most surprising result in this study was that using alignment-based conservation to identify regions enriched with TFBSs did not improve recall.
We finally report some potentially interesting cases that emerge from the analysis of the entire human kinome using alignment-free distances, and a brief analysis of the Entamoeba histolytica kinome.
To enhance the quality of the results, we filter away inconsistent elements using alignment-based techniques.
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