Custom iScale Oligos™

iScale Oligos DNA and siRNA are milligram and gram quantities of custom DNA and siRNA oligonucleotides manufactured to meet specialized needs.

DNA

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). For iScale quantities of NGS adapters, please see our Next-Gen Sequencing Oligos product

Product Benefits

• Amounts available to meet your experimental and commercial needs
• Consultation with our scientific team to set specifications for your application
• State-of-the-art analytical laboratory, ensuring a high-quality product

Product Features

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 dnaoligos@milliporesigma.com.

iScale Oligos DNA Specifications

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 Locked Nucleic Acid available 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

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

Quality Assurance

At the foundation of our manufacturing processes is a robust Quality Management System (QMS), which 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 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.

siRNA

Milligram and gram quantities of MISSION in-vivo-quality siRNA are superior for RNAi research in animals. We also manufacture other types of RNA, including miRNA and single-stranded RNA.

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

Product Features

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.

iScale Oligos siRNA Specifications

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 •  Locked Nucleic Acid available in every country outside of the United States •  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 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. Each replicate is a different mouse

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

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

References

1. Stany MP, Vathipadiekal V, Ozbun L, Stone RL, Mok SC, Xue H, Kagami T, Wang Y, McAlpine JN, Bowtell D, et al. Identification of Novel Therapeutic Targets in Microdissected Clear Cell Ovarian Cancers. PLoS ONE. 6(7):e21121. https://doi.org/10.1371/journal.pone.0021121

2. Beghin A, Belin S, Hage-Sleiman R, Brunet Manquat S, Goddard S, Tabone E, Jordheim LP, Treilleux I, Poupon M, Diaz J, et al. Correction: ADP Ribosylation Factor Like 2 (Arl2) Regulates Breast Tumor Aggressivity in Immunodeficient Mice. PLoS ONE. 4(11): https://doi.org/10.1371/annotation/b0d43779-c9aa-44fe-a46d-71d7d2bc4a6a

3. Ramachandran V, Arumugam T, Langley R, Hwang RF, Vivas-Mejia P, Sood AK, Lopez-Berestein G, Logsdon CD. The ADMR Receptor Mediates the Effects of Adrenomedullin on Pancreatic Cancer Cells and on Cells of the Tumor Microenvironment. PLoS ONE. 4(10):e7502. https://doi.org/10.1371/journal.pone.0007502

4. Brahmamdam P, Watanabe E, Unsinger J, Chang KC, Schierding W, Hoekzema AS, Zhou TT, McDonough JS, Holemon H, Heidel JD, et al. 2009. TARGETED DELIVERY OF siRNA TO CELL DEATH PROTEINS IN SEPSIS. 32(2):131-139. https://doi.org/10.1097/shk.0b013e318194bcee

5. Fitamant J, Guenebeaud C, Coissieux M, Guix C, Treilleux I, Scoazec J, Bachelot T, Bernet A, Mehlen P. 2008. Netrin-1 expression confers a selective advantage for tumor cell survival in metastatic breast cancer. Proceedings of the National Academy of Sciences. 105(12):4850-4855. https://doi.org/10.1073/pnas.0709810105

6. Watabe M, Aoyama K, Nakaki T. 2008. A Dominant Role of GTRAP3-18 in Neuronal Glutathione Synthesis. Journal of Neuroscience. 28(38):9404-9413. https://doi.org/10.1523/jneurosci.3351-08.2008