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Second, we used a 16S database of reference sequences trimmed to 100 nt centered on the V6 loop.
The 25 nt centered peak in D. melanogaster matched the previously reported piRNA peak of this species [ 6].
Further confirmation of this phenomenon was obtained through NMF analysis of three sequence sets, artificial sequences with and without inserted patterns and 8500 sequences with length 400 nt centered on putative human 3′-processing sites (Salisbury et al., 2006).
The nucleosome profiles of the [-600, 600] nt, centered on each ACS sequence, from the WT G1 sample were clustered using a k-mean clustering function with 10.000 iterations.
Second, flanking regions of each miRNA (500 nt centered on the miRNA) were used to search against the Rfam 10.0 library of covariance models (Griffiths-Jones et al. 2005) using Infernal 1.0 (Nawrocki et al. 2009) and Rfam-curated thresholds.
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The major peak, spanning from 20 to 25 nts and centered at 22 nt, correlated well with the size of mature miRNAs.
Motifs were predicted by searching for over-represented sequences within 100 nt windows centered on the TSSs.
For the production run, extract a 601 nt region centered on the SNP and exclude the SNP if this region contained an "N".
For VMS and VSS, extract the 121 nt region centered on the SNP and exclude the SNP if this region contained an "N".
Briefly, for each transcript the nucleotide with the greatest number of Pab1 hits was identified, and the height of this binding peak divided by the maximum height when reads in a 400 nt window centered on this peak were placed randomly.
Motif analysis (MEME, Bailey and Elkan, 1994) failed to identify an enriched RBM47-binding sequence in ±10 nt footprints centered on the top 3000 significant peaks (data not shown), but revealed a polyU sequence enriched in RBM47-binding sites containing cross-link induced mutations (deletion CIMS, Zhang and Darnell, 2011, Figure 4F).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com