uMRT Direct RNA-Seq Kit

Get longer reads, detect more genes and isoforms, and experience improved modification signal-to-noise from your direct RNA-Seq. The UltraMarathonRT® (uMRT) Direct RNA-Seq Kit pairs the helicase-driven processivity of uMRT with the Oxford Nanopore Technologies® (SQK-RNA004) workflow.

UltraMarathonRT delivers longer reads, higher yield, and superior isoform detection in Nanopore Direct RNA-Seq workflow.

Why Choose This Kit

• Detect more genes, isoforms, and longer, more contiguous native RNA reads
  High processivity at low temperature preserves RNA integrity and reduces early terminations and read truncation.

Direct RNA Sequencing of Universal Human Reference RNA using uMRT and Induro. After down-sampling to an equal number of total reads, the reads were aligned using minimap2, and transcriptome models were generated using IsoQuant followed by curation using SQUANTI3. The SQUANTI3 analysis confirms that uMRT identifies more isoforms per gene compared to Induro.

• Dramatically improved detection of long transcripts
  Retains long RNAs that regularly degrade at high temperatures (e.g., >50°C).This kit delivers >10% higher mapped reads from ≥ 2 kb RNAs. uMRT works at 30°C, which preserves the integrity of longer transcripts.

Number of mapped read counts per million (CPM) for read lengths ≥ 2 kb. Direct RNA sequencing performed on Human Brain Total RNA (Thermo, Cat#AM7962) using SQK-RNA004, N=2. All trials were down-sampled to 2.7M reads using SEQTK, then analyzed using NanoPlot.

• Generate Higher cDNA yields from your Sample
  Capture more of the sample. UltraMarathonRT’s optimized chemistry delivers higher cDNA yields for Direct RNA-Seq.
  

The uMRT Direct RNA-Seq kit generates higher library yields. Direct RNA Sequencing libraries of Universal Human Reference RNA and Human Brain RNA were created using uMRT and Induro. Results show that uMRT generates ~ 25% more RNA/cDNA yield from 1 µg of total RNA.

• Drop-in for ONT workflows
  Fits naturally into the Direct RNA-Seq protocol; no changes to downstream ligation, loading, or basecalling.
• Convenience & Cost-Savings
  One box contains everything you need to pair with ONT’s SQK-RNA004, eliminating the hassle and reducing the cost of ordering multiple reagents from different vendors.
  

Applications

• RNA modification mapping
• Isoform and UTR profiling directly on native RNA
• Viral and host–pathogen transcriptomics
• Long noncoding RNAs and repetitive/high-GC regions

UltraMarathonRT Direct RNA-Seq Kit Components:

• UltraMarathonRT® (20 U/μL)
• 2x RT Reaction Buffer
• dNTP Mix (10 mM each)
• RNA Conditioner
• 5x Ligation Buffer Q
• DNA Ligase (2,000,000 U/mL)
• RNase Inhibitor (40 U/μL)
• Quenching Buffer
• Nuclease-free Water

Reagents Required but Not Supplied:

• Oxford Nanopore Technologies® (ONT) SQK-RNA004 or SQK-RNA004-XL Direct RNA Sequencing kit.
• ONT MinION®/GridION® RNA Flow Cell (FLO-MIN004RA) or PromethION® RNA Flow Cell (FLO-PRO004RA).
• RNA Purification Beads.
• Qubit dsDNA HS Assay Kit (ThermoFisher, Q32851).
• RNA template: 500 ng - 1 μg total cellular RNA or 300 ng of poly (A)+ RNA is recommended for each reaction.

Product Information:

Catalog# R1008S (6 reactions), R1008M (48 reactions)

Storage Conditions: For short duration of storage (< 6 month), UltraMarathonRT can be stored in a -20℃ freezer. -80℃ is recommended for long-term storage. All other kit components are stable in a -20℃ freezer for long-term storage. Note that UltraMarathonRT can endure 10 freeze-thaw cycles without losing noticeable activity. After 20 freeze-thaw cycles, UltraMarathonRT maintains 90% of its original activity.

Downloads:

Brochure

UltraMarathonRT

2x RT Reaction Buffer

dNTP mix (10 mM each)

RNA Conditioner

5x Ligation Buffer Q

DNA Ligase

RNase Inhibitor

Quenching Buffer

Direct RNA-Seq Protocol

CoA

For Research Use Only.