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What is Boost?

Boost is a proprietary buffer that dramatically improves detection of target RNAs when RNA input is low. If your sample has < 10 ng of RNA, you should include it in your workflow. However, if your sample has > 10 ng of RNA, it is not necessary to include Boost in the workflow.

Can I buy Boost separately?

Boost is only available as part of an UltraMarathonRT kit.

What is the difference between MarathonRT & UltraMarathonRT?

UltraMarathonRT is a highly optimized version of MarathonRT. We mutated the enzyme to improve the overall efficiency of cDNA synthesis and the sensitivity to low abundance RNA detection, while retaining the ultra-high processivity of MarathonRT.

Is UltraMarathonRT as powerful as MarathonRT in opening stable RNA structures?

Yes, the helicase activity of UltraMarathonRT is the same as MarathonRT.

Can I replace MarathonRT with UltraMarathonRT in SHAPE-MaP and DMS-MaP?

UltraMarathonRT is an excellent choice for both SHAPE-MaP and DMS-MaP.

Can I order MarathonRT from RNAConnect?

If you require MarathonRT for your research, please contact us. While UltraMarathonRT performs substantially better than MarathonRT, we recognize that some workflows may still require MarathonRT. We have limited supplies available to accommodate such instances.

Can I order RT buffer separately?

Not at this time.

Will UltraMarathonRT work in the 10X Chromium system? Oxford Nanopore? PacBio?

UltraMarathonRT works well for most applications calling for cDNA synthesis. We have created and sequenced libraries constructed using UltraMarathonRT in Illumina NGS and Oxford Nanopore long-read sequencing platforms. We do not yet have data in the PacBio or Chromium systems. If you are interested in optimizing a workflow using UltraMarathonRT for these and other innovative platforms, please contact us to discuss the project with our technical team.

What are Group II Intron RTs?

Group II introns are ribozymes that catalyze their own excision from precursor RNAs and the subsequent ligation of flanking exons (a process known as self-splicing). This class of RTs, known as maturase RTs, contains an N-terminal extension (RT0) and a specific set of insertions between the seven conserved sequence motifs that are found in all RTs (RT1–7) (Blocker et al. 2005; Zhao and Pyle 2017). Maturase RT enzymes tend to be highly processive, as this is required for successful copying of the large, highly structured group II intron RNA and for successful propagation of group II introns within their hosts (Mohr et al. 2013; Lambowitz and Belfort 2015).

What are the optimal storage conditions and shelf life for UltraMarathonRT?

UltraMarathonRT can be stored at -20C for up to 3 months and should be stored at -80C for longer durations. When stored at -80C, uMRT maintains performance for >2 years.

Is UltraMarathonRT stable at 4°C?

UltraMarathonRT stability data at 4°C is coming soon.

Will UltraMarathonRT work at temperatures lower than 20°C? How long should I incubate it?

UltraMarathonRT can fully transcribe an 8kb PCR product at room temperature (20°C). Temperatures below 20°C have not been tested. UltraMarathonRT can copy an 8kb template in 10 min. We recommend a general incubation time of 15 minutes. Although longer times (up to 90 min) may be called for when working with very low abundance samples. Temperatures below 20°C have not been tested.

Will UltraMarathonRT work at temperatures higher than 42°C?

Please use UltraMarathonRT as directed in the protocols provided. UltraMarathonRT loses all activity above 45°C.

What is the longest incubation time I can use for UltraMarathonRT?

At 42°C, UltraMarathonRT has a transcription rate of 26 nt/second. UltraMarathonRT has been shown to actively transcribe for at least 90 minutes at both 42°C and room temperature. A maximum incubation time at varying temperatures has not been established.

Will UltraMarathonRT be inactivated after freeze-thaw?

UltraMarathonRT can endure 10 freeze-thaw cycles without losing noticeable activity. After 20 freeze-thaw cycles, UltraMarathonRT maintains 90% of its original activity.

Can I use any DNA polymerase with UltraMarathonRT?

Yes! Specific DNA polymerases should be chosen based on the experiment. For instance, we recommend a high-fidelity DNA polymerase such as KAPA HiFi for generating sequencing libraries and LongAmp Taq for targeted long cDNA amplification.

Do I need to purify the cDNA for the next-step of the experiment, e.g., PCR amplification?

cDNA purification prior to PCR amplification is optional. The actual requirement of cDNA purification depends on the amount of cDNA product and the DNA polymerase used in the PCR reaction.

When will additional kits (e.g. 1-step RT-PCR) become available?

Kits for RT-PCR applications will be available later this year. Additional Library Prep Kits will also be available near the end of 2024.

Which applications are best suited to UltraMarathonRT?

All applications where the production of cDNA is needed. Please contact our Technical Support Team if you are thinking of new, innovative applications for UltraMarathonRT.

What type of RNA samples work with UltraMarathonRT?

UltraMarathonRT works on any RNA samples ranging from total cellular RNA to targeted RNAs.

What is the dynamic range of input RNA?

UltraMarathonRT is sensitive to a wide range of RNA inputs. If your sample is between 10 ng -1pg, please use the Boost buffer provided in your UltraMarathonRT kit. If your sample is > 10 ng, you do not need to use Boost.

When will these kits be available in my region?

RNAConnect is building out our global distribution channels. Currently, we sell these kits directly to our customers in the USA and Canada and sell them through partners in the following countries: UK, Ireland, Germany, France, Italy, Spain, Portugal, Switzerland, Belgium, Austria, and Luxembourg. Please see our Distributors page (coming soon) for more information. If your region is currently not listed, please contact us.

Can I design a custom Template Switching Oligo to use with UltraMarathonRT?

You can design your own custom TSO, however, the RNAConnect proprietary TSO has been optimized to work best with our UltraMarathonRT Template Switching kit.

Can these kits be used in an automated workflow?

If you are interested in optimizing a workflow using UltraMarathonRT for these and other innovative platforms, please contact us to discuss the project with our technical team.