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By default, STAR outFilterMultimapNmax parameter (read alignments will be output only if the read maps fewer than this value) is set to 10. Should that just be set to 1 if you are going to process the data with htseq-count, which discards multi-mapped reads? Is there a benefit to leaving it at 10?
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Even if you keep multimappers in the SAM output, HTseq will not count them - it recognizes the unique mappers by NH:i:1 attribute.Originally posted by id0 View Post
Whether multimappers are useful depends on your downstream analysis. I find it useful to generate wiggle tracks separately for unique and multi-mappers, sometimes you can miss expression if you only look at unique mappers. Also, other quantification software (e.g. Cufflinks) can "rescue" multi-mappers. Typically multimappers do not occupy a lot of space, and - if needed - can be easily filtered by NH flag or the MAPQ field.
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Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...03-22-2024, 06:39 AM
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