I understand that we can either fractionate RNA or cDNA so poly A library is not a necessity for RNA-seq. However, would fractionated RNA alone serve to give a decent coverage?
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PolyA selection allows for selection of mRNA. Total RNA from a cell/tissue contains more than 98% of ribosomal RNA sequences and therefore it would be wise to do either polyA selection or remove ribosomal RNA by other means. If not done, you end up having 80% of your reads mappings to ribosomes.Post-doctoral Fellow
John Curtin School of Medical Research
Australian National University, Canberra, ACT, Australia
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