title: Synthesis and purification in a single column on a high-throughput automated oligonucleotide production system.
abstract: Oligonucleotides are synthesized and purified in a single column filled with a mixture of nucleoside-loaded and underivatized, high-cross link, non-swelling polystryene beads. A new oligonucleotide production system has been developed to completely automate-the entire process of sequence entry, phosphoramidite synthesis, cleavage, deprotection, purification, quantitation, and sample collection. Up to 48 primer-length, high-purity oligonucleotides can be produced in a 24 hour period of unattended automation on the ABI 3948 DNA Synthesizer. Synthesis, cleavage, and purification occur within the unique OneStepTM column. Analyses by MicroGel capillary electrophoresis, anion-exchange HPLC, PAGE, and enzymatic digestion/HPLC routinely exhibit > 90% product purity. Stringent PCR, automated fluorescent sequencing, and genetic analysis experiments show the positive effects of the total automation of purified oligonucleotide production.
authors.affiliation: Applied Biosystems Division of Perkin Elmer Co., Foster City, CA 94404, USA.
authors.forename: A
authors.initials: A
authors.lastname: Andrus
authors.forename: P
authors.initials: P
authors.lastname: Wright
authors.forename: J
authors.initials: J
authors.lastname: Wang
authors.forename: B
authors.initials: B
authors.lastname: Mullah
authors.forename: J
authors.initials: J
authors.lastname: Baier
authors.forename: J
authors.initials: J
authors.lastname: Mason
authors.forename: G
authors.initials: G
authors.lastname: Kaufman
mesh_terms: D001483:Base Sequence; D001671:Biochemistry / Q000295:instrumentation* / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D005069:Evaluation Studies as Topic; D007202:Indicators and Reagents; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction
chemical_list: D007202:Indicators and Reagents; D009841:Oligonucleotides