An Improved Method for Large Scale Synthesis of Oligonucleotides Applying the NPE/NPEOC-Strategy

Weiler, Jan; Pfleiderer, Wolfgang

doi:10.1080/15257779508012501

Cited by 9 publications

(6 citation statements)

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“…± The application of the normal oligonucleotide protocol to the phthaloyl-protected phosphoramidites required the checking of each step of the cycle regarding the stability of the new building blocks under the standard conditions. The acid-catalysed condensation and dimethoxytrityl-cleavage step, as well as the capping procedure, turned out to be compatible with earlier findings, whereas the oxidation procedure had to be changed from the normal I 2 treatment in pyridine/H 2 O/ THF to a treatment with 1m solution of tert-butyl hydroperoxide [30] in MeCN to avoid side reactions ( Table 1). A series of oligo-2'-deoxyribonucleotides ( Table 2) were synthesized in a machine-aided manner to give homogeneous products of high purity.…”

Section: Methodssupporting

confidence: 78%

Nucleotides, Part LXI, Phthaloyl Strategy: A New Concept of Oligonucleotide Synthesis

Beier

Pfleiderer

1999

HCA

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Section: Methodssupporting

confidence: 78%

Nucleotides, Part LXI, Phthaloyl Strategy: A New Concept of Oligonucleotide Synthesis

Beier

Pfleiderer

1999

HCA

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“…For the preparation of oligonucleotide hybrid arrays between the 3 -terminal nucleoside and the solid phase in combination with an alternative phosphoramidite that combine usage in hybridization screening experi- approach, the so-called ''NPE/NPEOC strategy'' (19), In Situ Oligonucleotide Synthesis which on CPG and polystyrene supports had led to oliInitially oligomer synthesis on the polypropylene gonucleotide products of high purity (20,21). This syn-sheets with the NPE/NPEOC nucleoside derivatives thesis strategy uses the b-eliminating base-protecting was carried out under running standard phosphoragroups NPE and NPEOC.…”

Section: Reagents O-[(ethoxycarbonylmentioning

confidence: 99%

Combining the Preparation of Oligonucleotide Arrays and Synthesis of High-Quality Primers

Weiler

Hoheisel

1996

Analytical Biochemistry

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purposes. For clinical applications like the antisenseBased on the oligomer-chip technology, oligonucleo-strategy (reviewed in Ref. 2), gram or even kilogram tide arrays were synthesized directly on polypropyl-amounts of oligonucleotides are required, prompting ene sheets by a modified phosphoramidite chemistry efforts on an up-scaling of synthesis yields (e.g., 3). For using b-eliminating nucleobase-protecting groups in many applications in molecular biology, on the other combination with a succinate solid-phase linker. This hand, notably DNA characterization and screening by method decouples the oligonucleotide deprotection hybridization and enzymatic assays like the polymerfrom the support cleavage procedure, in contrast to ase chain reaction (PCR) and DNA sequencing, oligostandard phosphoramidite chemistry. In addition to mer quantities in the picomole range are usually an being reliable substrates for hybridization experi-adequate amount, but larger numbers of oligonucleoments, the arrays also serve as source for the isolation tides are needed. Again, this has led to procedures aimof individual oligonucleotides. Technically, this al-ing at the simultaneous production of different oligonulowed for a direct control of the quality of the arrayed cleotides in small quantities (e.g., 4, 5). quence of an unknown nucleic acid is reconstructed from its hybridization binding pattern on a matrix which contains a comprehensive set of short oligonucleotide sequences (e.g., all 65,536 octamers). Initiated by the combination of solid-phase technolDuring our work concerned with developments toogy and the phosphoramidite chemistry introduced by ward a practical application of the SBH technique, such Beaucage and Caruthers (1), there has been much prog-as the ordering and selection of DNA fragments suitress in the automation of DNA synthesis. Today, the able for SBH analysis (10, 11), for example, or the sepreparation of synthetic oligonucleotides needed in bio-quence-independent leveling of oligomer binding stalogical, biomedical, and physical applications is usually bilities (12), it was apparent to us that such oligomer executed with commercial, automated DNA synthe-chips by design would be an ideal tool not only for sizers. These machines produce oligonucleotides in the screening procedures but also for the synthesis of nanomole up to micromole range. However, over the past few years, there have been two diametrical tend-2 Abbreviations

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“…3.1.8A to C) instead of a primary amino group (such as LCAA-CPG) creates linkages that are resistant to cleavage by the non-nucleophilic base DBU. These linkages have been used with base labile 5′-Fmoc-protecting groups to make acid-sensitive oligodeoxyribonucleotides and oligoribonucleotides (Brown et al, 1989) or with 2-(4-nitrophenyl)ethoxycarbonyl/NPE baseprotecting groups to allow on-column deprotection (Stengele and Pfleiderer, 1990;Weiler and Pfleiderer, 1995). A triethylamine-resistant sarcosine-succinic acid linker arm (Fig.…”

Section: Linker Arms For the Deprotection Of Immobilized Products Wit...mentioning

confidence: 99%

Solid‐Phase Supports for Oligonucleotide Synthesis

Pon

2000

CP Nucleic Acid Chemistry

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This unit begins with a discussion of the advantages and disadvantages of oligonucleotide synthesis using solid supports. The physical and chemical properties of solid-phase supports are discussed in terms of their suitability for oligonucleotide synthesis. In addition, the unit outlines the properties of linkers used for transient or permanent attachment of properly protected nucleosides to the derivatized support, as well as strategies for coupling nucleosides to linkers and conditions for the release of synthetic oligonucleotides from specific supports.

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An Improved Method for Large Scale Synthesis of Oligonucleotides Applying the NPE/NPEOC-Strategy

Cited by 9 publications

References 0 publications

Nucleotides, Part LXI, Phthaloyl Strategy: A New Concept of Oligonucleotide Synthesis

Nucleotides, Part LXI, Phthaloyl Strategy: A New Concept of Oligonucleotide Synthesis

Combining the Preparation of Oligonucleotide Arrays and Synthesis of High-Quality Primers

Solid‐Phase Supports for Oligonucleotide Synthesis

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