Exact(1)
Raw reads were first filtered to discard low-quality reads and FastQC (Babraham Bioinformatics) used to inspect read base qualities.
Similar(59)
Methylation rates were estimated by inspecting read sequence and recording the location of CpGs relative to the reference genome.
Our study has also shown the benefit of computing inter-sample SNP-concordances and inspecting read alignments in order to attain more confident results.
Furthermore, we carefully inspected reads originating from tRNAs and snoRNAs, since recent reports have highlighted a novel class of small RNAs originating from these structural RNAs [ 8- 12].
IGV version 2.3 was used to visually inspect the read alignments for selected samples [ 10].
FastQC was used to inspect the reads for quality issues before proceeding with the assembly.
IGV (http://www.broadinstitute.org/igv/) was used for manually inspect the reads assembled to each gene.
For all 5 events we manually inspected the read pileups in IGV to look for possible artifacts.
Inspecting the read sequences derived from mature miRNA and miRNA* pairs, we found that the 5' ends were significantly less variable than the 3' ends.
However, Artemis can be used to visualize and confirm this error by inspecting the read and mapping qualities and BAM flags of the alignments.
We visually inspected individual read alignments from the F1 father for the remaining 400 loci and eliminated a further 81 SNPs (18.3%) from contigs that we assumed included paralogs, that is, contigs with three or more haplotypes per individual (no single SNP position had more than two alleles, but there were multiple haplotypes within the contig of a single individual).
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