Details

Autor(en) / Beteiligte

• Habus, Ivan
• Agrawal, Sudhir

Titel

Improvement in the synthesis of oligonucleotides of extended length by modification of detritylation step

Ist Teil von

• Nucleic acids research, 1994-10, Vol.22 (20), p.4350-4351

Ort / Verlag

England: Oxford University Press

Erscheinungsjahr

1994

Quelle

MEDLINE

Beschreibungen/Notizen

• 4,4'-Dimethoxytrityl, (DMTr), is at present commonly used for the protection of the 5'-terminal hydroxy function in solid phase DNA or RNA synthesis. During the oligonucleotide synthesis the first step, prior to coupling with the appropriately protected phosphoroamidite monomer, is the removal of the 5'-protecting group, (DMTr, detritylation) by a suitable protic acid. We have found that dichloroacetic acid in the presence of 0.1% of anhydrous alcohol, (e.g., methanol or ethanol), or a freshly distilled 1H-pyrrole is a highly effective non-depurinating detritylating agent for oligonucleotide synthesis on a solid support. The assembly of oligonucleotides was performed on a 0.75 or 1.0 mu mole scale using CPG-1000 or -2000 and beta -cyanoethyl phosphoramidites on automated DNA synthesizer, (Milligen/Bioresearch 8700 series). The standard program cycles were used, except detritylation time was extended from 68.5 sec. to 308.5 sec., followed by wash extended from 10 sec. to 130 sec. All syntheses were repeated at least three times, under the same conditions for each sequence. The crude products, after release from the support were analysed by gel-capillary electrophoresis.