Automated Purification and Quantification of Oligonucleotides. (October 1999)

• Record Type: Journal Article
• Authors: Ivanetich, K.M.; Reid, R.C.; Ellison, R.; Perry, K.; Taylor, R.; Reschenberg, M.; Mainieri, A.; Zhu, D.; Argo, J.; Cass, D.; Strickland, C.
• Is Part Of: Biotechniques. Volume 27:Number 4(1999)
• Page Start: 810
• Page End: 823
• Publication Date: 1999-10
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/99274rr04
• Languages: English
• ISSNs: 0736-6205

Abstract

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.