PCR GENERATION OF LARGE AMOUNTS OF PURIFIED DNA

The preparation and purification of PCR generated DNA fragments suitab le for footprinting and classical drug binding studies is described. O ne of the fragments, a 214-mer derived from pBR322 DNA exhibits a biph asic melting profile. This behavior appears to be due to a non-random distribution of base pairs within the fragment causing a region rich i n AT base pairs to melt prior to a segment having a high concentration of GC base pairs. The usefulness of large amounts of PCR generated DN A for footprinting and optical binding studies involving drugs is also presented and discussed.