RiboCop for Human/Mouse/Rat plus Globin

Total RNA from mammalian blood is comprised of highly abundant undesired RNA species, such as ribosomal RNA (rRNA), accounting for ~80 – 90 % of total RNA, and globin mRNA, representing 30 -80 % of all mRNAs. Lexogen’s RiboCop rRNA Depletion Kit for Human/Mouse/Rat plus Globin removes undesired cytoplasmic (28S, 18S, 5.8S, and 45S rRNA) and mitochondrial rRNA (mt16S, mt12S, and 5S), and globin mRNA from intact as well as degraded whole blood RNA.

RiboCop for Human/Mouse/Rat plus Globin (HMR+Globin) is offered as a stand-alone kit (Cat. No. 145) or as a bundled version with the CORALL Total RNA-Seq Library Prep Kit (Cat. No. 147). RiboCop Kits are compatible with all random primed total RNA-Seq library prep kits.

Globin Depletion Increases Gene Detection

RiboCop for Human/Mouse/Rat plus Globin (HMR+Globin) simultaneously depletes rRNA and globin mRNA, providing a highly convenient workflow for blood samples and freeing up sequencing space for RNAs of interest (Fig. 1). Thus, removal of globin mRNA leads to a significant increase in gene detection (Fig. 2).

Fig_01 - RiboCop HMR plus Globin

Figure 1 | RiboCop rRNA Depletion for Human/Mouse/ Rat plus Globin (HMR+Globin) efficiently removes rRNA and globin mRNA from human blood RNA. A) RNA was extracted from fresh human donor blood using SPLIT RNA Extraction. 10 ng whole blood RNA were depleted with RiboCop HMR V2 or HMR+Globin. Libraries were prepared with the CORALL Total RNA-Seq Library Prep Kit and sequenced on NextSeq500 (1×75 bp). Reads were mapped against the human reference genome (GRCh38.95) and reads mapping to rRNA (blue) and globin (purple) were calculated. B) Correlation plot for depleted blood samples shown in A (uniquely mapping reads). Major globins (marked in red) are retained in samples depleted with HMR V2 and efficiently removed by RiboCop HMR+Globin depletion.

Fig_02 - RiboCop HMR plus Globin

Figure 2 | Increased gene detection upon globin depletion. A) The number of detected genes per number of reads uniquely mapping to exons (FeatureCounts) was plotted for the samples shown in Figure 1. B) Bar plot illustrating the number of detected genes for the samples shown in Figure 1 at 1 M reads /sample, normalized to Counts Per Million (CPM) at a threshold of CPM > 1.

RiboCop Performance Across Species

RiboCop enriches RNAs of interest by efficiently removing rRNA and globin mRNA from human, mouse, and rat whole blood RNA (Fig. 3).

Fig_03 - RiboCop HMR plus Globin

Figure 3 | RiboCop rRNA Depletion for Human/Mouse/ Rat plus Globin (HMR+Globin) enriches for RNAs of interest in blood sample across species. 100 ng whole blood RNA were depleted with either RiboCop HMR V2 or RiboCop HMR+Globin. Library preparation, sequencing, and data analysis was performed as described in Figure 1. Reads were mapped against the respective reference genomes for human (GRCh38.95), mouse (mmu_GRCm38.95), and rat (rno_Rnor_6.0.95), respectively.