Two common forms of the human MLH1 gene may be associated with functional differences

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Abstract
Genomic DNA was purified using standard phenol-chloroform extraction methods and amplified for 30-35 cycles in 20 μl (3-5 pmol of each primer, 50 μmol/l of each dNTP and 0.1 unit ofExtra-pol II DNA polymerase from Eurobio, Les Ulis, France). Primers and cycling conditions used to amplify microsatellites were as described in the Genome Database,http://gdbwww.gdb.org. The upstream primer was labelled at its 5′ end using IRD800 labelling (Lincoln, NE). A total of 0.5-1 μl of reaction product was denatured and electrophoresed on denaturing polyacrylamide gels (obtained by mixing 19 ml of 8% Sequagel XR from National Diagnostics, Atlanta, GA, with 952 μl of Long Ranger, FMC, Rockland, ME). Samples were separated and analysed on a Li-Cor 4000 automated sequencer.