DNA and siRNA Custom iScale Oligos™

Section Overview

• Why are scalable oligos useful?
• Scale Oligos DNA Specifications
• Commercial and cGMP Oligo Capabilities
• siRNA
• iScale Oligos siRNA Specifications
• Demonstrated Use in vivo
• Related Products

WHY ARE SCALABLE OLIGOS USEFUL?

Scalable oligos are crucial because they allow for flexible production in varying quantities to meet specific research or commercial needs. This scalability ensures efficient resource utilization, cost-effectiveness, and adaptability to project demands. Whether it's small-scale experiments or large-scale commercial applications, scalable oligos offer the versatility and convenience required for streamlined workflows and optimized outcomes in research and applied genomics.

iScale Oligos DNA and siRNA offer scalable production of milligram to gram quantities of custom DNA and siRNA oligonucleotides. This scalable manufacturing capability caters to specialized needs in commercial applications, ensuring efficient and cost-effective production of high-quality oligos for research, diagnostics, lab developed tests (LDTs) and therapeutic development.

Scalable Custom DNA Oligos

With 35+ years expertise in oligo manufacturing, we are a leading supplier of scalable custom oligonucleotides globally. Milligram and gram quantities of Custom DNA Oligos are for in vivo, high-throughput, and commercial projects (life science research tools, molecular diagnostics, and laboratory developed tests). iScale custom oligos are batch validated under stringent QMS control to meet unique customer needs.

Custom DNA oligo benefits

• Oligonucleotide quantities available to meet your experimental and commercial needs with fast and efficient global manufacturing scalability
• Direct consultation with our scientific team to set specifications for your application
• State-of-the-art analytical laboratory, ensuring a tightly-controlled, high-quality product

Scalable custom oligo DNA specifications

iScale oligos are available in a wide range of sequence lengths, quantities, purifications and formats. For anything marked as ‘Inquire’ below or if you have custom oligo needs that are different from the general specifications presented, please send a request to dnaoligos@milliporesigma.com.

Quantities	10, 25, 50, 100, 250 & 500 mg  1, 2, 5, 10, & 15 g (inquire for larger quantities)
Purification	Desalt, RP-HPLC & in vivo Treatment
Sequence Lengths	10 to 55 bases (inquire for longer lengths)
Modifications	6-FAM, Biotin, Phosphate, Amino-Modifiers & others  Inquire for feasibility of Locked Nucleic Acid & other modifications
Quality Control	100% mass spectrometry  Analytical HPLC available   Certificate of Analysis available
Format	Supplied dry or in solution in tubes or plates  Pooling, aliquoting & normalization available (inquire for feasibility)
Services	Salt Exchange: replaces toxic ammonium ions from the synthesis with physiologically-tolerable sodium ions. Included for all purification types.    Filtration: minimizes bacteria, molds and other contaminants. Included for in vivo treatment purification.    Endotoxin testing: ensures gram-negative LPS is below an acceptable threshold, otherwise the host may die from toxic shock as initiated by and immune cascade. Available for in vivo treatment purification.

Useful Applications for scalable custom oligos

• Antisense
• Binding studies
• High-throughput PCR/qPCR
• Sanger Sequencing & NGS (non-adapters)
• X-ray crystallographic structure determination
• Immunostimulatory assays
• Microarray production

Custom oligo manufacturing Quality Assurance

At the foundation of our oligo manufacturing processes is a robust Quality Management System (QMS) that drives compliance to our following Quality Registrations:

• ISO 9001:2015 for manufacturing research-grade oligonucleotides
• ISO 13485:2016 for manufacturing diagnostic-grade oligonucleotides
• ISO 14001:2015 for effective environmental management

This quality culture is integrated into all aspects of our custom oligo business. Key components of our QMS include:

• Certificates of analysis
• Document control
• Change notification
• Vendor management
• Business agreements
• Corrective and preventive actions

Each component drives quality assurance, and for verification, we routinely host rigorous registrar, internal, and customer audits.

Commercial and cGMP Oligo Capabilities

As part of our Commercial Oligos offering, our Custom cGMP oligos come with consistency and the highest quality. Whether you want to commercialize or develop an assay around qPCR, isothermal amplification, NGS, or other technology, our products strike the right balance between breadth and depth.

Scalable Small Interfering RNA (siRNA)

Our global manufacturing expertise ensures milligram and gram quantities of MISSION^® in vivo quality siRNA, ideal for superior RNAi research in animals. Additionally, we produce other RNA types like miRNA and single-stranded RNA, all scalable to meet diverse research needs with excellence.

siRNA Product Benefits

• Suitable for in vivo applications, including target validation and pre-clinical testing
• Available for MISSION Predesigned siRNA or your custom siRNA sequences
• Appropriate for direct delivery or as an essential component of your formulation strategy

iScale Oligos siRNA Product Specifications

Custom scalable siRNA are available in a range of quantities, purifications and formats depending on your manufacturing needs. For anything marked as ‘Inquire’ below or if you have needs that are different from the other general specifications presented, please send a request to sirnarequest@milliporesigma.com.

Quantities	•  3, 50 & 100 mg •  Inquire for larger quantities
Purification	• Desalt, RP-HPLC & in vivo
Sequence Form	• 19 to 50mer simplexes or duplexes
Modifications	•  Biotin, phosphate, Amino-Modifier (C3, C6, C6 dT, C7, C12), 6-FAM™, Cyanine 3,    Cyanine 5, Cyanine 5.5, 2'-OMe & phosphorothioate •  Inquire for feasibility of Locked Nucleic Acid & other modifications
Quality Control	• 100% mass spectrometry* • Analytical HPLC available • Certificate of Analysis available
Format	• Supplied dry in tubes or plates • Pooling & aliquoting available (inquire for feasibility)
Services	• If the oligonucleotide is to be used in vivo, in vivo grade purification and these services   are recommended:   - Salt exchange (replaces toxic ammonium ions from the synthesis     with physiologically-tolerable sodium ions)   - Filtration (minimizes bacteria, molds, and other contaminants)   - Endotoxin testing (ensures gram-negative LPS is below an acceptable     threshold, otherwise the host may die from toxic shock as initiated by     an immune cascade)

*Depending on manufacturing site, PAGE may be used to assess siRNA duplexes.

Demonstrated use of precision siRNA in vivo

We teamed up with Bioo Scientific (now PerkinElmer) to validate our MISSION in vivo quality siRNA (see article in Drug Discovery & Development—Aug 1, 2008).

In these experiments, H1155luc cells were injected into NOD/SCID mice. Tumors formed and were injected with luciferase-specific siRNA or non-targeting in vivo quality siRNA formulated in MaxSuppressor™ in vivo RNA-LANCEr II. After 24 hours, D-luciferin solution was injected, and the animals were imaged using the NightOWL II LB 983 instrument from Berthold Technologies. Animals treated with luciferase-specific siRNA showed significant reduction of luciferase activity while non-targeting siRNA-treated animals were positive for luminescence (Figure 1). These data demonstrate the effectiveness of in vivo quality siRNA formulated in MaxSuppressor™ in vivo RNA-LANCEr II in vivo delivery agent.

Figure 1. Luciferase-specific siRNA treated animal on the left; non-targeting siRNA treated animal on the right

At 72 hours post-injection, the tumors were removed, and the protein was extracted and normalized using a Bradford assay. Luciferase concentrations were assayed using a Luciferase ELISA. Protein reduction is represented relative to a non-targeting RNAi agent treated animal. Luciferase activity was reduced by two-fold in the tumors (Figure 2).

Figure 2. Percent luciferase protein expression in Luciferase-specific siRNA treated animals and controls measured by ELISA. Each replicate is a different mouse.

If additional help is needed, please consult our technical services group at oligotechserv@milliporesigma.com. 

MISSION is a trademark of Merck KGaA, Darmstadt, Germany and/or its affiliates. Label License.

References

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