MASSEUNDERSOEKELSESMETODE FOR DETEKSJON AV ENKELTBASEMUTASJONER I MULTIPLE LOCI I GENOMISK DNA VED BRUK AV DENATURERINGSGRADIENT-GELELEKTROFORESE FULGT AV EFFEKTIV OVERFOERINGOG HYBRIDISERING TIL FORSKJELLIGE PROBER

摘要

PCT No. PCT/NO88/00074 Sec. 371 Date May 31, 1989 Sec. 102(e) Date May 31, 1989 PCT Filed Oct. 3, 1988 PCT Pub. No. WO89/02930 PCT Pub. Date Apr. 6, 1989.Single base pair differences between otherwise identical DNA fragments can result in an altering of the melting behavior which can be detected by denaturing gradient gelelectrophoresis (DGGE). A method has been developed for efficient transfer of genomic DNA fragments from the gel following DGGE. The DGGE is run in a polyacrylamide gel (PAG) containing 2% low gelling temperature agarose (LGT). The PAG is crosslinked with a reversible crosslinker, and after electrophoresis the crosslinks are cleaved and 80-100% of the DNA fragments are transferred to nylon membranes by alkaline transfer. Hybridization with gene specific probes is then performed. The technique has been used to identify an RFLP in the COLIA2 gene.