Recently we have performed RNA sequencing using Ribo-Zero in order to remove rRNA from fragmented FFPE samples. The number of reads passing filter are around 30 million per sample. The samples were mapped using Tophat and the readcounts were calculated using HTSeq-count.There aren't much reads left that map to the ribosomal RNA, which is a good thing, but the number of reads that were assigned to a gene according HTSeq-count were not as high as we hoped to see (3-12 million reads per sample).
It seems that there are many reads that do not map to unique locations and a lot of unmapped reads do map to human (BAC) DNA clones which results in a loss of >60% of the reads that were passing filter.
However, the reads that are assigned to do show a very nice profile (compared to DSN-treatment, which we used to use).
Does anyone experience the same?
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Metagenomics has improved the way researchers study microorganisms across diverse environments. Historically, studying microorganisms relied on culturing them in the lab, a method that limits the investigation of many species since most are unculturable1. Metagenomics overcomes these issues by allowing the study of microorganisms regardless of their ability to be cultured or the environments they inhabit. Over time, the field has evolved, especially with the advent...-
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