A method for partial digestion of total human DNA with restriction enzymes has been developed on the basis of a principle already utilized by P. A. Whittaker and E. Southern (1986, Gene 41: 129-134) for the analysis of phage .lambda. recombinants. Total human DNA irradiated with uv light of 254 nm is partially digested by restriction enzymes that recognize sequences containing adjacent thymidines because of TT dimer formation. The products resulting from partial digestion of specific genomic regions are detected in Southern blots by genomic-unique DNA probes with high reproducibility. This procedure is rapid and simple to perform because the same conditions of uv irradiation are used for different enzymes and probes. It is shown that restriction site polymorphisms occurring in the genomic regions analyzed are recognized by the "allelic" partial digest pattterns they determine.
PARTIAL DIGESTION WITH RESTRICTION ENZYMES OF UV-IRRADIATED HUMAN GENOMIC DNA A METHOD FOR IDENTIFYING RESTRICTION SITE POLYMORPHISMS
NOBILE C;
1988
Abstract
A method for partial digestion of total human DNA with restriction enzymes has been developed on the basis of a principle already utilized by P. A. Whittaker and E. Southern (1986, Gene 41: 129-134) for the analysis of phage .lambda. recombinants. Total human DNA irradiated with uv light of 254 nm is partially digested by restriction enzymes that recognize sequences containing adjacent thymidines because of TT dimer formation. The products resulting from partial digestion of specific genomic regions are detected in Southern blots by genomic-unique DNA probes with high reproducibility. This procedure is rapid and simple to perform because the same conditions of uv irradiation are used for different enzymes and probes. It is shown that restriction site polymorphisms occurring in the genomic regions analyzed are recognized by the "allelic" partial digest pattterns they determine.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
