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The sequence must not contain any in-frame stop codons or BspQI recognition sites. Sequences may contain either a PaqCI or a BsaI recognition site, but not both. Stop codons are selected separately from the drop-down menu.

We recommend a dual stop codon sequence (e.g., TGATAG) to promote efficient translation termination. However, any single or dual stop codon combination may be selected.

Our standard offering includes an optimized, IP-free Human β-globin UTR set containing a Kozak sequence and designed for high-throughput synthesis workflows. If custom UTRs are preferred, please provide the desired 5′ and 3′ UTR sequences.

For compatibility with co-transcriptional capping, the 5′ UTR must include the appropriate +1 and +2 nucleotides immediately downstream of the promoter sequence. For most mRNA applications, these nucleotides are typically AG. For enzymatic capping, which requires a custom workflow, we strongly recommend GG for potent T7 polymerase activity. These bases immediately follow the promoter sequence (e.g., TAATACGACTCACTATA for T7 RNA polymerase). All custom 5′ sequences will be reviewed for compatibility.

To clearly define the start of the poly(A) tail, the 3′ UTR must not end with an adenosine (A) residue. Stop codons and poly(A) tail lengths are selected separately from the drop-down menus and should not be included in the submitted 3′ UTR sequence.

Our standard offerings include either a continuous 100A poly(A) tail or a segmented 30/70A poly(A) tail design. These formats are suitable for most mRNA applications and represent commonly used tail architectures. For projects based on published or proprietary constructs, custom poly(A) tail lengths and designs can be incorporated to match the original sequence requirements.

Enter the custom poly(A) tail sequence for your construct. This sequence must contain only DNA bases (A, T, C, G).

For custom products, the minimum production scale for each product depends on the number of custom products in your order:

• 4 or more custom products: minimum 0.1 mg per product.
• 2–3 custom products: minimum 0.5 mg per product.
• 1 custom product: minimum 1 mg.

These minimums apply only to custom products. Catalogue products have no minimum order requirement.

These minimum order requirements allow us to offer small-scale production at significantly reduced cost. For production scales below 1 mg, labour and operational costs substantially outweigh material costs. By establishing minimum order quantities, fixed project costs can be distributed across multiple products, resulting in favourable pricing models compared to individual low-scale orders.

We can source and incorporate a broad range of modified nucleotides to meet project-specific requirements. Our standard formulation uses 100% replacement of UTP with N1-methylpseudouridine triphosphate (N1mΨTP). Common alternatives include fully canonical nucleotides, Cy5-labelled UTP (5% or 20% substitution of total UTP), and 100% replacement of CTP with 5-methyl-CTP.

Cy5-labelled transcripts offer fluorescent signal for visualization and tracking applications, although higher levels of Cy5 incorporation may reduce RNA translational efficiency. We can provide guidance on selecting an appropriate labelling level based on your experimental objectives.

LiCl: We routinely perform an optimized LiCl precipitation process for research-grade mRNA production at scales up to 10 mg. This method provides a flexible and cost-effective purification option, with good selectivity for long ssRNA over dsRNA, DNA, proteins, and short RNA fragments. However, LiCl precipitation is not GMP-compatible and is generally less amenable to scale-up than chromatography-based purification approaches.

Silica Column: We offer silica column purification for production scales of 1 mg and below. This method is well suited to small-scale purification and utilizes workflows commonly employed in research laboratories.

Oligo(dT) Chromatography: We maintain a range of Sartorius BIA Separations™ oligo(dT) monolith columns, from spin-column formats through semi-preparative scale, enabling purification across a wide range of project sizes. All purification work is performed on dedicated RNA-only chromatography systems, including Vanquish HPLC and ÄKTA FPLC platforms. Our optimized binding and elution methods are designed to maximize target recovery while improving target-to-off-target capture ratios.

Our standard process uses co-transcriptional Cap 1 capping with a GAG-compatible cap analog, providing an efficient and cost-effective approach for most mRNA applications. For projects requiring alternative initiation sequences, enzymatic capping workflows, or Cap 0, Vaccinia and Faustovirus capping systems are available upon request.

We offer both wild-type T7 and engineered low-dsRNA T7 polymerases. While wild-type T7 remains a robust and economical choice for most research applications, the low-dsRNA variant reduces double-stranded RNA impurities generated during transcription and is recommended when higher transcript purity is required. This is particularly important for non-vaccine applications, or where dsRNA impurities may influence downstream biological activity, innate immune responses, or experimental outcomes.

To support long-term storage and minimize freeze-thaw cycles, products ordered at 1 mg or greater are aliquoted into 1 mg vials by default, unless otherwise requested. For orders below 1 mg, the full requested amount is provided in a single vial. Alternative aliquoting configurations are available upon request. Additional charges apply for large aliquoting needs.

We offer high-resolution RNA integrity analysis using a SCIEX MDQ-Plus capillary electrophoresis system with optimized separation conditions. This analysis provides an electropherogram and integrated peak data that can be used to estimate full-length transcript purity and identify degradation products. While CE can also be used for size estimation, migration can vary as capillaries age and are replaced, reducing sizing reproducibility over time. For this reason, we generally recommend gel electrophoresis for transcript size assessment. However, CE-based sizing can be provided when required.

We offer quantitative dsRNA analysis using optimized slot blot assays. Note that as dsRNA measurements are highly dependent on assay design, standards, and calibration methods, results are often not directly comparable between laboratories.

The High-Throughput option is designed for efficient screening of multiple samples and intra-blot comparisons.

Inter-blot variability can approach 20%, making side-by-side analysis within the same blot the preferred approach for direct sample comparisons.

We offer quantitative dsRNA analysis using optimized slot blot assays. Note that as dsRNA measurements are highly dependent on assay design, standards, and calibration methods, results are often not directly comparable between laboratories.

The High-Precision option utilizes four dilutions per sample to ensure measurements are collected within the optimal assay dynamic range, improving precision and comparability between experiments.

Inter-blot variability can approach 20%, making side-by-side analysis within the same blot the preferred approach for direct sample comparisons.

Poly(A) tail length analysis is performed through an optimized multi-RNase digestion workflow that selectively degrades non-poly(A) regions, followed by capillary electrophoresis characterization of the remaining poly(A) homopolymers or LC-MS. This service is currently offered as an Early Access assay while standards and validation procedures continue to be refined.

We have CE and LC-MS capabilities for assessing mRNA capping efficiency; however, method development is ongoing to evaluate robustness and reproducibility across different transcript architectures and UTR sequences. As a result, capping analysis is currently available on a custom-project basis. Please contact us if capping characterization is a critical project requirement.

Cellulose purification enriches ssRNA via selective adsorption of dsRNA. We use an optimized and scalable method based on the study found here.