RT-qPCR

Providing Accurate CRISPR KO/KD Results

The UltraMarathonRT® (uMRT) Two-Step RT-qPCR Kit provides reliable, sensitive quantitation of the RNAs in your sample. The kit is powered by UltraMarathonRT which is a novel RT with intrinsic helicase activity and strand displacement ability that enables high fidelity cDNA from any RNA regardless of length, structure or sequence. Conventional RT-qPCR systems use retroviral reverse transcriptases (eg MMLVs) that systematically create artifacts across RNA-targeting CRISPR platforms (Csm, PspCas13b, CasRx) — meaning the data they produce are unreliable.

The cause: guide RNA co-purifies with cellular RNA and blocks MMLV reverse transcriptases from reading through the guide-binding region, making “no knockdown” look like knockdown. The uMRT Two-Step RT-qPCR Kit doesn't have this problem. Its robust strand displacement activity reads straight through bound guide RNA, so you can use the spanning-amplicon designs you already prefer and get accurate, reliable KO/KD values every time.

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Reliable, Precise, and Timely 

The UltraMarathonRT Two-Step RT-qPCR Kit ensures unmatched data reliability by providing consistent Ct values regardless of primer type, RNA complexity, or GC content, eliminating biases and ensuring accurate quantification. With high inhibitor tolerance and a streamlined protocol that delivers results in under an hour, this versatile kit is ideal for RNA-seq validation, small RNA quantification, alternative splicing analysis, and is the only RT-qPCR kit that entirely solves the KO/KD artifact issue created by many CRISPR system gRNAs.

Key Features

Resolves CRISPR Artifacts

Provides consistent Ct values regardless of amplicon position relative to the guide RNA binding site. Conventional RTs give you a different apparent knockdown efficiency depending on where your primers land — the uMRT Two-Step RT-qPCR Kit gives you the same, accurate answer every time.

Adapted from Watkins et al.

Provides consistent Ct value regardless of primer type (e.g. gene specific, oligo(dT) or randomer) or location. Other RT-qPCR kits will provide widely different Ct values if primers are in high vs low GC content areas, whereas the uMRT Two-Step RT-qPCR Kit will provide consistent Ct values regardless of location.

Data Reliability

Robust Performance

Reads through protein-bound and guide-RNA-bound RNA that stalls conventional retroviral RTs. This is the mechanism behind the Johns Hopkins finding: uMRT's intrinsic helicase clears the guide-binding region instead of stopping at it, eliminating the false-positive knockdown signal seen with MMLV-based enzymes.

Consistent Ct values are maintained in the presence of inhibitory compounds potentially found in biological sample preparation such as ethanol (resistant to 10%), isopropanol (resistant to 5%), TRIzol (resistant to 0.7%), Formalin (resistant to 0.0025%), Ammonium Acetate (resistant to 50mM), Bile Salts (resistant to 0.05%), and Hematin resistant to 10µM).

High Inhibitor Tolerance

Minimal Experimental Time

An optimized protocol allows researchers to generate complete RT-qPCR data in under 1 hour.

UltraMarathonRT has intrinsic helicase activity for RNA structure unwinding, making the high temperature reaction conditions utilized by other RTs totally unnecessary.

Preserves RNA Template Integrity

Dye-Based Detection

Incorporates reliable dye-based technology to facilitate accurate and reproducible real-time monitoring of RNA levels.

The reverse transcriptase used in the Johns Hopkins study that identified and resolved the guide-RNA RT-qPCR artifact. If you're running RNA-targeting CRISPR (Csm, PspCas13b, CasRx) or other guide-based knockdown systems, this is the kit built to give you a result you can trust.

Validated for CRISPR Knockdown Confirmation

Built for researchers running RNA-targeting CRISPR knockdown experiments who need RT-qPCR results free of KO/KD artifacts — and for anyone robust, reliable data.

Learn More about uMRT for RT-qPCR