Seamless Whole Transcriptome Solutions for Blood
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Whole Transcriptome RNA-Seq for Blood
Get the best sequencing results from whole blood samples with Lexogen’s fast and flexible whole transcriptome solutions for Blood! CORALL for Blood Whole Transcriptome RNA-Seq provides high sensitivity, optimal coverage and globin removal regardless of RNA quality.
The CORALL for Blood Whole Transcriptome RNA-Seq bundle enables fast and cost-efficient generation of stranded, UMI-labelled, and unique dual indexed libraries with adjustable insert size. The streamlined workflow including simultaneous rRNA and globin mRNA depletion and library preparation is completed in under 6 hours – compatible with RNA input from 1 ng – 1000 ng whole blood RNA, including low quality RNA (RIN <3)!
- Transcript expression analysis
- Isoforms or splicing studies
- Detection of fusion transcripts
- Analysis of long non-coding RNAs
- snoRNA and snRNAs detection
- circRNA analysis
The workflow is compatible with various blood collection tubes, including PAXgene, ETDA tubes, and other systems.
Performance
Total RNA from mammalian blood is comprised of highly abundant undesired RNA species. Ribosomal RNA (rRNA) accounts for ~80 – 90 % of total RNA and globin mRNA represents 30 – 80 % of all mRNAs. Lexogen’s RiboCop rRNA Depletion Kit for Human/Mouse/Rat plus Globin combined with CORALL RNA-Seq efficiently generates NGS libraries devoid of 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.
Validated Performance on Whole Blood RNA
Depletion of rRNA and globin mRNA frees up sequencing space for RNAs of interest (Fig. 1). This workflow allows to study coding and non-coding RNAs of the blood, including mRNA, lncRNAs, snoRNAs, and snRNAs.
Figure 1 | CORALL for Blood with simultaneous depletion efficiently removes rRNA and globin mRNA from human whole blood RNA. RNA was extracted from human donor whole blood. 5 ng whole blood RNA were depleted with RiboCop HMR V2 or HMR+Globin. Libraries were prepared with the CORALL Total RNA-Seq V2 Library Prep 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 (red) were calculated.
Increased Gene Detection from Whole Blood Samples
Removal of rRNA and globin mRNA prior to CORALL library preparation significantly increases gene detection from whole blood samples (Fig. 2).
Whole Blood mRNA-Seq Workflow to Efficiently Increase mRNA Reads
The combination of CORALL mRNA-Seq with an additional RiboCop depletion step for removal of rRNA and globin mRNA provides a convenient workflow to process also stored blood samples in only 7 hours (Fig. 3). This workflow frees up sequencing space for mRNAs of interest (Fig. 4) and leads to a significant increase in gene detection.
Figure 3 | CORALL mRNA-Seq V2 with an additional rRNA and globin depletion step efficiently increases usable reads for mRNA-Seq applications using whole blood samples. Lexogen RNA-Seq workflows for blood samples are compatible with established blood collection systems, including PAXgene.
Figure 4 | CORALL for Blood mRNA-Seq combines rRNA and globin depletion with mRNA selection enabling elaborate mRNA-Seq applications, such as splice site detection and isoform analysis from whole blood samples.
“We have evaluated a double depletion method along with poly(A) capture using the CORALL RNA-Seq kit on isolated RNA from human blood collected in PAXgene tubes. When comparing combined rRNA and haemoglobin depletion (RiboCop rRNA Depletion Kit HMR plus Globin) with only using poly(A) capture (Poly(A) RNA selection kit), we saw less multimapping of reads. In addition, we managed to identify additional expressed genes whilst seeing a strong correlation between expression levels in genes identified in both sets. We were very pleased with this improvement. The workflow of using poly(A) selection combined with RiboCop HMR plus Globin and CORALL RNA-Seq library preparation was smooth and user-friendly and is highly recommended from a lab perspective.“
Elina Ekedahl, NGS Specialist, TATAA Biocenter
Workflow
FAQ
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Whole Transcriptome RNA-Seq for Blood
Safety Data Sheet
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Ordering Information
| Cat. No. | Product Name |
| 185.24 | RiboCop (HMR+Globin) and CORALL Total RNA-Seq V2 Library Prep Kit with UDI 12 nt Set A1, 24 preps |
| 185.96 | RiboCop (HMR+Globin) and CORALL Total RNA-Seq V2 Library Prep Kit with UDI 12 nt Set A1, 96 preps |
| 186.24 | RiboCop (HMR+Globin) and CORALL Total RNA-Seq V2 Library Prep Kit with UDI 12 nt Set B1, 24 preps |
| 186.96 | RiboCop (HMR+Globin) and CORALL Total RNA-Seq V2 Library Prep Kit with UDI 12 nt Set B1, 96 preps |
| Associated Product | |
| 157.96 | Poly(A) RNA Selection Kit V1.5, 96 preps |
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