Automated Purification and Quantification of Oligonucleotides | Zendy

Kathryn M. Ivanetich | Zendy; Ralph Reid | Zendy; Ross Ellison | Zendy; Kathy M. Perry | Zendy; R.G. Taylor | Zendy; M. Reschenberg | Zendy; Avantika Mainieri | Zendy; Debin Zhu | Zendy; J. Argo | Zendy; Darrell L. Cass | Zendy; Corey Strickland | Zendy

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Automated Purification and Quantification of Oligonucleotides

Author(s) -

Kathryn M. Ivanetich,

Ralph Reid,

Ross Ellison,

Kathy M. Perry,

R.G. Taylor,

M. Reschenberg,

Avantika Mainieri,

Debin Zhu,

J. Argo,

Darrell L. Cass,

Corey Strickland

Publication year - 1999

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/99274rr04

Subject(s) - oligonucleotide , yield (engineering) , chromatography , cartridge , extraction (chemistry) , dna , chemistry , materials science , biochemistry , metallurgy

We have developed automated methods for the trityl-on purification and quantification of synthetic oligonucleotides. Oligonucleotide purification is by solid-phase extraction cartridges using Amberchrom CG-50 resin on an XYZ-axis robotic system. Quantification is by OD260nm using an online UV-visible spectrophotometer with sipper. The purification of 20 oligonucleotides requires 5 min of user set-up time, plus 20 min per sample of robot time. For a 15-25-mer at the 40 nmol scale of synthesis, the method gives a yield of 2.8 ODs from a load of 10.1 OD, i.e., a 28% average yield. Oligonucleotides purified by this method have proven to be successful for primers for automated DNA sequencing.

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