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As common aligners can allow a small amount of mismatches during alignment, reads containing SNPs may still be mapped to the reference.
The initial alignment reads and their re-mapped tails composed "piece-alignments", which were then clustered if similarly mapped tails were shared based on location and orientation.
Specifically, for each sample, we mapped the filtered BWA alignment reads to a chromosome and exact base pair locations for input into the CNV-seq software.
Prior to alignment, reads were trimmed using a sliding window of 8nt.
Before alignment, reads with a low quality and adapters were detected using FastQC and removed.
The RNA2MAP mapping tool generated two types of alignment: reads uniquely mapped to miRNAs and reads generating multiple hits.
Similar(49)
To make TSS alignments, aligned reads were counted using genomeCoverageBed for every base pair.
Once CCS reads are sorted, the pipeline uses Bowtie2's default alignment (end-to-end read alignment) to align reads to the corresponding reference sequence and outputs a set of alignments in SAM format.
Duplicate alignments (reads aligning to the same genomic position) were removed using SAMtools [ 13].
Initial visual inspection of sequence alignments indicated that the Ribo-Zero libraries showed more 'background' alignments (reads aligned in intergenic and intronic regions) than the poly(A) datasets.
Reads that corresponded to flag 1796 in the bam alignment file (flag 1796: read unmapped, not primary alignment, read fail quality check, read is PCR or optical duplicate) were filtered out.
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