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@sial.com.

 

iScale Oligos DNA Specifications
Quantities • 10, 25, 50, 100, 250 & 500 mg
• 1 to 5 g (inquire for larger quantities)
Purification • Desalt (≤5 g), RP-HPLC (≤1 g) & In Vivo
Sequence Lengths • 10 to 50 bases (inquire for longer lengths)
Modifications • Biotin, Biotin TEG, Biotin dT, phosphate, Amino-Modifier (C3, C6, C6 dT, C7, C12),
  TET™,  HEX™, 5-Me-dC, Spacer (12, 18, C3), dSpacer, Thiol-Modifier-C3 S-S,   
  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 or in solution in tubes or plates
• Pooling, aliquoting & normalization 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
   


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@sial.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.

 

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

 

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).
 

Each replicate is a different mouse


Figure 2
– Each replicate is a different mouse
 

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

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

 

Materials

     

References

siRNA

  • Stany MP, Vathipadiekal V, Ozbun L, Stone RL, Mok SC, Xue H, Kagami T, Wang Y, McAlpine JN, Bowtell D, Gout PW, Miller DM, Gilks CB, Huntsman DG, Ellard SL, Wang YZ, Vivas-Mejia P, Lopez-Berestein G, Sood AK & Birrer MJ (2011). Identification of novel therapeutic targets in microdissected clear cell ovarian cancers. PLoS One, 6, e21121.
  •  Beghin A, Belin S, Hage-Sleiman R, Brunet Manquat S, Goddard S, Tabone E, Jordheim LP, Treilleux I, Poupon MF, Diaz JJ & Dumontet C (2009). ADP ribosylation factor like 2 (Arl2) regulates breast tumor aggressivity in immunodeficient mice. PLoS One, 4, e7478.
  • Ramachandran V, Arumugam T, Langley R, Hwang RF, Vivas-Mejia P, Sood AK, Lopez-Berestein G & Logsdon CD (2009). The ADMR receptor mediates the effects of adrenomedullin on pancreatic cancer cells and on cells of the tumor microenvironment. PLoS One, 4, e7502.
  • Brahmamdam P, Watanabe E, Unsinger J, Chang KC, Schierding W, Hoekzema AS, Zhou TT, McDonough JS, Holemon H, Heidel JD, Coopersmith CM, McDunn JE & Hotchkiss RS (2009). Targeted delivery of siRNA to cell death proteins in sepsis. Shock, 32, 131-9.
  • Fitamant J, Guenebeaud C, Coissieux MM, Guix C, Treilleux I, Scoazec JY, Bachelot T, Bernet A & Mehlen P (2008). Netrin-1 expression confers a selective advantage for tumor cell survival in metastatic breast cancer. Proc Natl Acad Sci U S A., 105, 4850-5.
  • Watabe M, Aoyama K & Nakaki T (2008). A dominant role of GTRAP3-18 in neuronal glutathione synthesis. J Neurosci, 28, 9404-13.