An optimized PCP-based procedure for production of C-13/N-15-labeled DNA

We have substantially improved a procedure that we previously described for producing C-13/N-15-labeled DNA (Chen et ah, FEES Left. 436, 372-376, 1998 ) to provide an economical and straightforward approach to the preparation of labeled DNA. The conditions for the PCR reactions have been optimized to permit the use of low concentrations of the costly labeled dNTPs (50 muM f or each). In addition, a rapid and high-yield purification procedure has be en developed that allows us to obtain a high yield of very pure labeled DNA . These modifications to our original procedure permit us to obtain 1.9 mg of an 18 bp DNA oligomer from 20 nag of dNTPs (ca. 10% yield from the start ing dNTPs). This is sufficient material for the preparation of 0.4 mM sampl e in a volume of 400 mul. In summary, this procedure is a cost-effective, t ime-efficient procedure for the production of labeled DNA for NMR studies. (C) 2001 Academic Press.