Tackling Transcriptomics with New Line of RNase H-based Depletion Reagents
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New England Biolabs has announced an expansion of its RNA depletion product portfolio to support more sensitive and cost-effective RNA sequencing via next-generation sequencing platforms. The NEBNext RNase H-based RNA depletion protocol offers efficient and specific depletion of the abundant RNA species that interfere with the analysis of both coding and non-coding RNAs of biological significance. The newly-released NEBNext Globin & rRNA Depletion Kit (Human/Mouse/Rat) and NEBNext rRNA Depletion Kit (Bacteria) complement the existing NEBNext rRNA Depletion Kit (Human/Mouse/Rat). All kits are available both with and without RNA Sample Purification Beads, and have a 2-hour workflow, requiring less than 10 minutes of hands-on time. The kits also support a broad range of input amounts of total RNA, from low ng to 1 µg, and are effective with intact or degraded RNA.
The NEBNext Globin & rRNA Depletion Kit depletes adult, fetal and embryonic globin mRNA (HBA1/2, HBB, HBD, HBM, HBG1/2, HBE1, HBQ1 and HBZ), as well as cytoplasmic rRNA (5S, 5.8S, 18S, 28S, ITS and ETS) and mitochondrial rRNA (12S and 16S). "As researchers who work with blood samples know, globin mRNAs – together with ribosomal RNAs – are the most abundant RNA species present within blood. Removing these is essential to the efficient sequencing and analysis of the less-prevalent RNAs," said Eileen Dimalanta, Ph.D., Associate Director of Applications & Product Development at NEB. "NEB's RNase H-based depletion method makes it possible to efficiently and specifically deplete even poor-quality RNA."
The NEBNext rRNA Depletion Kit (Bacteria) removes the abundant bacterial rRNAs (5S, 16S, and 23S) resulting in the enrichment of bacterial mRNAs and non-coding RNAs of interest. This method is targeted to both gram-positive and gram-negative organisms and is effective with RNA from monocultures or mixed bacterial species, such as those used in metatranscriptome studies. "Bacterial mRNA's lack of a poly(A) tail reduces the options for isolation of biologically relevant RNA species for cost-effective analysis. This new kit addresses the inability to use oligo d(T)-based methods to enrich mRNAs of interest from bacterial samples, by instead efficiently removing the more-abundant rRNAs, across a range of bacterial species," stated Fiona Stewart, Ph.D., Portfolio Marketing Manager for NEBNext. "By taking the dominant rRNAs out of the equation, it becomes easier to see the true picture."
These new NEBNext RNA depletion kits complement the NEBNext products for RNA library preparation, including the Ultra™ II Directional RNA Library Prep Kit for Illumina® and the NEBNext Library Quant Kit for Illumina, as well as the Monarch® Total RNA Miniprep Kit.