Biomolecular Resource Laboratory

Location:
Room 109
Bowman Gray Technical Center
Building 611-13W
WFUBMC
Medical Center Blvd.
Winston-Salem, NC 27157

Shipping Address:
950 Reynolds Blvd.
Building 611-12 Dock 1
Attn: WFU Building 611-13W
Winston-Salem, NC 27105

Director:
Mark O. Lively, Ph.D.,
Professor of Biochemistry

Technicians:
Gulya Kourman
Mark Morris

Phone: (336) 716-6138

Fax: (336) 716-7200

Email: dnasynth@wfubmc.edu

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The DNA synthesis core lab provides custom-made quality oligonucleotides to assist you in your research. The oligos are synthesized on an automated Applied Biosystems Inc. Model 394 DNA/RNA synthesizer. This model was chosen because of its high coupling efficiency and reliability. The synthesis begins with the 3' terminal nucleoside that is attached to a solid support and the nucleotide bases are added one at a time until the synthesis is complete.

Each machine-grade oligo is cleaved and deprotected, precipitated by addition of ethanol to remove salt, residual synthesis reagents and very short truncated nucleotide products. The quantity of synthetic product is determined by measuring the absorbance at A260 nm and then lyophilized before delivery.

This basic purification method is sufficient for routine PCR, sequencing, and probe hybridization assays. It is recommended however that oligos containing more than 40 base pairs be purified using a reverse phase cartridge (RPC). Longer primers contain more failure sequences and RPC purification helps remove most of these truncated products. We also suggest that at least the 200nmol scale be used for longer oligos that will be purified by RPC. RPC purification might be desired for cloning, protein binding assays, site directed mutagenesis or for any "special oligo" containing modified nucleotides (please see Special Oligo section).