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. 2014 May 27;5:157. doi: 10.3389/fgene.2014.00157

Figure 1.

Figure 1

Examples of Bioanalyser assay interpretation for a variety of RNAs. (A) Standard Eukaryotic RNA shows a 28S rRNA band at 4.5 kb that should be twice the intensity of the 18S rRNA band at 1.9 kb (human) resulting in a RIN = 8.0–10.0. Small peaks are sometimes present after the marker that represent 5S and 5.8S subunits, tRNAs and small RNA fragments about 100 bp; these are more obvious when using phenol or trizol exterection methods, QIagen columns will generally remove small RNAs. When degraded 28S RNA is reduced and more fragments are detected around the 18S RNA subunit resulting in RIN = 6.4, which is below the quality required for high throughput DNA sequencing. Invertebrate RNA results in fragmentation of the 28S rRNA into two bands that co-migrate with the 18S rRNA resulting in aberrant RIN score of <8.0 although the mRNA is unaffected and suitable for sequencing. Genomic DNA can skew the 28S RNA peak but can easily be remedied by RNAse-free DNase1 digestion. (B) Ribosomal RNA removal by isolation of poly-A-RNA assessed by Bioanalyser RNA assay.