sparQ RNA-Seq HMR Kit
- High quality directional RNA library prep ready to sequence in a single day
- Simple workflow with 3 reaction tubes, 9 steps and 10 components
- Efficient and integrated removal of rRNA and globin mRNA from
human/mouse/rat (HMR) samples - Improved results for samples with limited quantity and/or poor quality RNA
Now with Bead Booster:
- Shorter total time investment of 4.5 hours for over 35% savings in time*
- Up to 5x increase in yield*
(*) compared to typical NGS RNA-seq with ribo-globin depletion
sparQ RNA-Seq HMR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.
sparQ RNA-Seq HMR Kit
Description
sparQ RNA-Seq HMR Kit simultaneously depletes rRNA and globin mRNA while generating stranded RNA-seq libraries for Illumina® NGS platforms in 4.5 hours. In a single step and tube, RNA fragmentation and depletion of abundant ribosomal and globin transcripts (human, mouse, and rat) are integrated. The proprietary, highly optimized enzymes and streamlined workflow generate high quality, directional transcriptome NGS libraries from either intact or degraded RNA samples, with key improvements for low input and FFPE samples.
sparQ UDI Adapter Resources
Setting up sparQ Library Prep kits in Illumina Experiment Manager
Details
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Volume |
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Component Description |
Cap Color |
95216-024 |
95216-096 |
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Box 1 |
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Frag Prime RG Depletion Mix |
Blue |
1 x 96 μl |
1 x 384 μl |
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1st Strand Enzyme Mix |
Brown |
1 x 96 μl |
1 x 384 μl |
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2nd Strand Buffer |
Green |
1 x 480 μl |
2 x 960 μl |
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2nd Strand Enzyme Mix |
Green |
1 x 240 μl |
1 x 960 μl |
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Rapid Ligation Buffer (5X) |
Orange |
1 x 480 μl |
2 x 960 μl |
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T4 DNA Ligase |
Orange |
1 x 240 μl |
1 x 960 μl |
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HiFi Plus Master Mix (2X) |
White |
1 x 600 μl |
2 x 1.2 ml |
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Primer Mix |
White |
1 x 36 μl |
1 x 144 μl |
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Bag 2 |
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Bead Booster |
Clear |
1 x 144 μl |
1 x 576 μl |
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UDI Dilution Buffer |
Clear |
1 x 1.44 ml |
1 x 5.76 ml |
Performance Data
Streamlined workflow
sparQ RNA-Seq HMR Kit workflow is simplified to 3 reaction tubes, 9 steps and 9 components. rRNA and globin mRNA removal is integrated with the RNA fragmentation and priming step, enabling faster time to result, less hands-on time and fewer pipetting steps.
Efficient removal of rRNA
High proportion of reads mapped to protein coding regions. Libraries were prepared from various sample types at varying input amounts, then sequenced. Reads were mapped to the human transcriptome RNA biotypes. Libraries prepared with sparQ RNA-Seq HMR Kit showed the highest proportion of ‘protein coding reads, when compared with other supplier kits, and very low rRNA reads (0-5%).
Effective removal of globin mRNA
Consistent, high percentage of globin mRNA depletion. sparQ RNA-Seq HMR libraries were generated with 300 ng and 10 ng input of blood samples according to the standard protocol, then compared against a library generated without globin mRNA depletion at 300 ng input. Globin mRNA HBA1, HBA2 and HBG2 levels were measured using TPM (transcripts per million) count calculated with CLC Genomic Workbench 20.0.4. Abundant globin mRNA was efficiently depleted, allowing for data focused on mRNA and other RNA biotypes of high interest.
Higher yield
Higher library yield observed with sparQ RNA-Seq HMR Kit for low input and degraded input RNA. Comparable library yields were obtained with each kit using 1 µg of Universal Human Reference (UHR) RNA. For low input quantities of UHR RNA, the lowest recommended RNA input amount for each kit was used. A The sparQ RNA-Seq HMR Kit generated consistently higher yields at low UHR RNA inputs and B higher yields with low quality FFPE RNA (RIN 3.5) for both input amounts.
Better overall coverage uniformity
Uniform 3’ transcript coverage. sparQ RNA-Seq HMR Kit was uniquely able to retain uniform 3’ coverage for FFPE RNA, a feature that will help correctly identify full-length genes in low quality samples. For UHR RNA, all RNA-seq kits showed comparable uniformity.
Increased unique transcript identification
Comparison of unique fragments. The sparQ RNA-Seq HMR Kit consistently demonstrated higher rates of unique fragments indicating the highest library diversity regardless of RNA input quantity and sample which will enable more accurate quantification of low-level or rare transcripts and better transcript quantification.
Excellent data concordance
High data concordance between high vs. low RNA input amounts of various sample types was achieved. The libraries were sequenced on the Illumina® NextSeq550 instrument at 2 x 100 bp. The sequenced reads were analyzed by aligning the reads to the reference genome of each sample accordingly using the CLC Genomics Workbench 20.0.4 software (Qiagen®).
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sparQ RNA-Seq HMR Kit
Easy protocol and configuration. I haven’t compared the results with other products yet but the data itself came out pretty decent. Love it!
Times on hand was shorter than our current kit, and obtained quality was comparable. Therefore, we decided to implement this kit in our lab from now on.