Synthesis of Modified Antisense Oligonucleotides
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Corresponding Organization : University of Massachusetts Chan Medical School
Other organizations : Institute for Neurodegenerative Disorders
Protocol cited in 11 other protocols
Variable analysis
- Oligonucleotide synthesis method (MerMade 12 or AKTA Oligopilot 100)
- Solid support used for oligonucleotide synthesis (controlled pore glass functionalized with long-chain alkyl amine and Unylinker® terminus for unconjugated antisense strands, modified solid support for divalent sense strands)
- Oligonucleotide properties and quality (not explicitly mentioned)
- Phosphoramidite concentration (0.15M for MerMade 12, 0.2M for AKTA)
- Activator (5-(Benzylthio)-1H-tetrazole at 0.25M)
- Detritylation reagents (3% trichloroacetic acid in dichloromethane for MerMade 12, 3% DCA in toluene for AKTA Oligopilot)
- Capping reagents (CAP A: 20% NMI in ACN, CAP B: 20% Ac2O, 30% 2,6-lutidine in ACN)
- Sulfurization reagent (0.1 M solution of DDTT in Pyridine for 3 minutes)
- Phosphoramidite coupling time (8 minutes for all amidites)
- Not explicitly mentioned
- Not explicitly mentioned
Annotations
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