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The shorter forward read contains a barcode that can be used to map the read back to the spatial position of the microarray as well as an UMI to be able to account for PCR duplicates.
Then, SCISSORS attempts to map the read to known insertions.
Alignment candidate threshold (minimal alignment length to map the read) was optimized with hash size 20.
We used the suffix array based program Vmatch [ 22] to map the read sequences to the genome requiring a minimum of 90% identity over the full length alignment.
"Hash size" (k-mer length for alignment) and "alignment candidate threshold" (minimal alignment length to map the read) parameters were pre-optimized prior to the analysis with respect to alignment sensitivity and time needed for the analysis.
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BSMAP and Bismark were used to map the reads onto the reference genome.
BWA aligner was used to map the reads against the cassava genome.
Tophat2 (version 2.0.8) was applied to tissue samples to map the reads to the reference genome.
TopHat v2.0.6 [ 33] was used to map the reads to the Bos taurus UMD3.1 reference genome.
SOAPaligner/soap2 (http://soap.genomics.org.cn/) was used to map the reads to the assembled transcriptome.
As with many RNA-Seq analyses, the first step of IsoEM is to map the reads.
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