Consistent Temperature. Consistent Results.
The need for fast, field-deployable molecular assays has never been greater. Recombinase Polymerase Amplification (RPA) is a highly sensitive isothermal amplification method that enables rapid, onsite pathogen testing, including for infectious diseases. However, enzyme and protein lot-to-lot variability has historically limited its reliability - until now.
Key Features and Benefits
- Amplify consistently with individually QC'd enzymes and proteins, improving performance and reproducibility
- Overcome LAMP limitations with lower amplification temperatures, simple primer design, and shorter amplification times
- Go mobile with glycerol-free, lyophilization-friendly formulations, enabling robust field-based testing
Figure 1. Molecular representation of Recombinase Polymerase Amplification (RPA). T4 UvsX recombinase (green ovals) and UvsY protein (purple hexagons) bind to amplification primers, forming a complex that is complementary to sequences in double stranded DNA. T4 gp32 protein (pink dots) acts as a single-stranded binding protein and stabilizes the unwound DNA strand, allowing Bsu DNA Polymerase - Large Fragment (red ovals) to initiate strand displacing amplification. The repetition of the cycle leads to exponential amplification. Schematic recreated from James, et al, Diagnostics 2020, 10, 399.
Proteins Available
| Protein | Details |
| T4 UvsX Recombinase (glycerol -) | Quality controlled for enzyme activity (forming D-loop recombination structures for initiation of amplification) and freeze/thaw stable |
| T4 UvsY Protein (glycerol -) | Accessory protein to T4 UvsX Recombinase with robust single stranded binding |
| T4 Gene 32 Protein (glycerol +/-) | Stabilizes ssDNA to increase amplification efficiency and primer specificity |
| Bsu DNA Polymerase, Large Fragment (glycerol +/-) | Displays strong strand displacement and enables isothermal amplification at lower temperatures than Bst polymerase |
Be the first to know
Sign up for updates on the upcoming launch of Watchmaker's RPA enzymes, including product availability, technical details, and early access opportunities.
Key Performance Data
Access more information from FFPE samples
Processing FFPE samples is inherently challenging due to the template damage incurred during fixation and the presence of residual cross-links. Watchmaker's novel FFPE treatment step, paired with a reverse transcriptase specifically engineered for RNA-seq applications, delivers excellent sensitivity and enables researchers to address a broader range of clinically relevant samples effectively.
Access more information from FFPE samples
Increase sensitivity with ultra-low-input samples
Robust and reproducible sample processing can be difficult for applications and samples where RNA quantities are restrictive, such as fine needle biopsies, limiting the sensitivity of associated assays. Watchmaker RNA Library Prep Kits with Polaris Depletion enable the generation of high quality libraries with as little as 1 ng of RNA.
Increase sensitivity with ultra-low-input samples
Retain quantitative gene expression information across input amounts
Agreement between high- and low-input libraries with respect to unique genes identified and their abundance is indicative of how well sample complexity is maintained as RNA input amounts decrease. Ideally, the relative gene expression profile should remain consistent across input amounts for a given sample. Watchmaker better preserves the gene expression profile as RNA input amount decreases from 500 ng to 10 ng, whereas other commercial solutions result in data distortion with low inputs.
Figure 4. Prevent data distortion with... MORE
Retain quantitative gene expression information across input amounts
Figure 4. Prevent data distortion with low inputs. Libraries were prepared from a whole blood sample in triplicate using 500 ng and 10 ng of RNA and rRNA and globin depletion upstream of library prep. Data were randomly downsampled to 16M paired reads per library. Differential expression analysis between averaged 500 ng (control) and 10 ng whole blood samples using DESeq2. Results indicate that other vendors lose representation of low abundance genes at 10 ng, while the Watchmaker solution does not.
Efficiently sequence blood samples while covering lncRNAs
Overabundant globin mRNAs in blood samples pose a challenge for efficient RNA sequencing, as ≥ 30% of bases map to these transcripts if traditional mRNA capture is used. Additionally, mRNA capture omits a significant subset of long non-coding RNAs (lncRNAs) that have demonstrated regulatory functions.¹ Polaris Depletion removes both rRNA and globin mRNA while also providing excellent coverage of lncRNAs, such as MALAT1 which has been implicated in at least 17 cancer types, including lung, breast, and pancreatic.²
- Statello, L., Guo, CJ., Chen, LL. et al. Gene regulation by long non-coding RNAs and its biological functions. Nat Rev Mol Cell Biol 22, 96–118 (2021).
- Amodio, N., Raimondi, L., Juli, G. et al. MALAT1: a druggable long non-coding RNA for targeted anti-cancer approaches. J Hematol Oncol 11, 63 (2018).
Efficiently sequence blood samples while covering lncRNAs
| RNA input range | 1 - 1000 ng |
|---|---|
| Polaris Depletion Kit - rRNA/Globin (HMR) contents | Depletion Master Mix Depletion Probes - rRNA/Globin (HMR) Probe Digestion Master Mix |
| Watchmaker RNA Library Prep Kit contents | FFPE Treatment Buffer Frag & Prime Buffer 1st Strand Buffer 1st Strand Enzyme 2nd Strand Enzyme 2nd Strand Buffer Ligation Enzyme Ligation Buffer Equinox® Library Amplification Master Mix (2X) P5/P7 Primer Mix (10X) |
| Shipping conditions | Ice packs |
| Storage | -20°C ± 5°C |
| Shelf life | (24 rxn): ≥ 6 months (> 24 rxn): ≥ 12 months |
Can the Watchmaker RNA Library Prep Kit with Polaris Depletion be used to detect coding RNAs?
This kit depletes total RNA of ribosomal and globin RNA. Both coding RNA and long non-coding RNA are represented in the sequencing data.
To focus on only coding RNA, please use the Watchmaker mRNA Library Prep Kit. for coding RNA.
What other RNA Library Prep Kits does Watchmaker offer?
The Watchmaker mRNA Library Prep Kit includes poly(A) enrichment to focus sequencing on coding RNA. This kit is compatible with intact RNA samples.
The Watchmaker RNA Library Prep Kit is suitable for total RNA and typically used upstream of target enrichment. This kit is compatible with both high quality and degraded RNA samples.
Can this kit be automated?
The Watchmaker RNA Library Prep Kit with Polaris Depletion is highly automatable. The kit was developed with automation in mind and 96 reaction kits include generous overages to account for dead volumes on liquid handling platforms. Visit our automation page for a list of scripts already developed or to contact a member of our automation team regarding your platform and needs.
Is this kit compatible with other (non-Illumina) sequencing platforms?
This kit has been demonstrated to be compatible with Element, Singular and Ultima sequencing platforms.
Does this product support long-read sequencing?
We use random primers to prime during first strand synthesis, which does not support full-length cDNA generation. This makes it unlikely to be compatible with-long read applications.
Do you offer custom formats?
Yes! Watchmaker offers customer fills, packaging, concentrations, and labeling - including private label - designed to meet your unique needs. We offer flexible terms to serve organizations of any size, and our right-sized processes enable rapid turnaround time on customization. Please contact sales@watchmakergenomics.com to learn more about our capabilities.
Are you ISO 13485-certified?
Yes! Our Quality Management System has achieved ISO 13485:2016 Certification. Our certificate has been awarded for the design, development, manufacture, contract manufacture, and support of high performing reagents for genomics applications in medical research. Download the certificate here.
| Description | 24 rxn | 96 rxn | |
|---|---|---|---|
| Watchmaker RNA Library Prep Kit with Polaris Depletion incl. reagents for rRNA and globin depletion, RNA library prep, and amplification | BK0002-024 | BK0002-096 | Request a quote |
Please contact sales@watchmakergenomics.com to inquire about custom kit configurations.
Validate your login