EFFICIENT SYNTHESIS OF P-32 LABELED SINGLE-STRANDED-DNA PROBES USING LINEAR PCR - APPLICATION OF THE METHOD FOR ANALYSIS OF STRAND-SPECIFICDNA-REPAIR

We have developed a rapid method to synthesize radioactively labeled s ingle-stranded DNA probes suitable for strand-specific analysis of sin gle copy genes on Southern blot. Linear PCR with 10 mu Ci alpha(32)P-d ATP (3000 Ci/mmol) as the only dATP source enabled us to generate stra nd-specific DNA probes with high specific activity. The probes synthes ized by this method have higher specific activities and the same stran d specificity compared to the end-labeled single-stranded DNA probes o btained from single-stranded M13mp18/19 vectors. Application of the me thod for strand-specific analysis of ultraviolet-induced DNA lesions i n defined DNA sequences significantly improved the hybridization signa l.