The performance of the GeneRead Pure mRNA Kit and mRNA purification kits from other suppliers was compared. Starting material for mRNA purification was 0.5 μg total RNA. Despite only requiring one purification round, the GeneRead Pure mRNA Kit provides superior enrichment of mRNA. Residual rRNA is still visible in the form of two distinct bands in samples prepared using products from other suppliers.
For analysis, 5 μg total RNA from Jurkat cells was treated using the GeneRead Pure mRNA protocol. Subsequently, libraries of the enriched samples, as well as of the total RNA were made using the same method and sequenced in multiplex by Illumina sequencing on a MiSeq system. Reads were mapped against the Biomart database and the percentage of mapped reads from each library for the biotypes was plotted. rRNAs are often listed under low qual. unmapped, as the annotation status of rRNA transcripts is often still suboptimal.