Preparation of 13C/15N-labeled oligomers using the polymerase chain reaction

摘要

Preparation of 13C/15N-labeled DNA oligomers using the polymerase chain reaction (PCR). A PCR based method for uniform (13C/15N)-labeling of DNA duplexes is described. Multiple copies of a blunt-ended duplex are cloned into a plasmid, each copy containing the sequence of interest and restriction Hinc II sequences at both the 5' and 3' ends. PCR using bi-directional primers and uniformly 13C/15N-labeled dNTP precursors generates labeled DNA duplexes containing multiple copies of the sequence of interest. Twenty-four cycles of PCR, followed by restriction and purification, gave the uniformly 13C/15N-labeled duplex sequence with a 30% yield. Such labeled duplexes find significant applications in multinuclear magnetic resonance spectroscopy.