explain secondary/supplementary aln for RNA-seq
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README.md
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README.md
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@ -154,6 +154,20 @@ slight improvement to accuracy. For noisy Nanopore Direct RNA-seq reads, it is
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recommended to use a smaller k-mer size for increased sensitivity to the first
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recommended to use a smaller k-mer size for increased sensitivity to the first
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or the last exons.
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or the last exons.
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Minimap2 rates an alignment by the score of the max-scoring sub-segment,
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*excluding* introns, and marks the best alignment as primary in SAM. When a
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spliced gene also has unspliced pseudogenes, minimap2 does not intentionally
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prefer spliced alignment, though in practice it more often marks the spliced
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alignment as the primary. By default, minimap2 outputs up to five secondary
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alignments (i.e. likely pseudogenes in the context of RNA-seq mapping). This
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can be tuned with option **-N**.
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For long RNA-seq reads, minimap2 may produce chimeric alignments potentially
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caused by gene fusions/structural variations or by an intron longer than the
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max intron length **-G** (200k by default). For now, it is not recommended to
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apply an excessively large **-G** as this slows down minimap2 and sometimes
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leads to false alignments.
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It is worth noting that by default `-x splice` prefers GT[A/G]..[C/T]AG
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It is worth noting that by default `-x splice` prefers GT[A/G]..[C/T]AG
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over GT[C/T]..[A/G]AG, and then over other splicing signals. Considering
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over GT[C/T]..[A/G]AG, and then over other splicing signals. Considering
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one additional base improves the junction accuracy for noisy reads, but
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one additional base improves the junction accuracy for noisy reads, but
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