terminator cycle sequencing kit on an abi 373 fluorescent sequencer abi foster city calif restriction digestion was performed following the pcr with tsp45 i according to manufacturer 39 s protocol new england biolabs beverly ma the digested pcr products were electrophoresed on a 6 visigel stratagene la jolla ca and visualized by ethidium bromide staining pedigree structure in fig2 has been slightly modified in order to protect patient confidentiality total rna was extracted from the lymphoblastoid cell line of an affected individual and first strand synthesis was performed by oligo dt priming gibco brl gaithersburg md primers 1f 5 u2032 acgacagtgtggtgtaaagg 3 u2032 and 13r 5 u2032 aacatctgtcagcagatctc 3 u2032 seq id no 10 corresponding to nucleotides 21 u2013 40 and 520 u2013 501 of genbank l08850 were used to amplify a product of 500 bp containing the mutation at nucleotide 209 pcr products were subjected to restriction digestion by tsp45 i the nucleotide at nt 209 creates a novel tsp45 i site fig1 so that the normal allele will be restricted in 4 fragments of 249 218 24 and 9 bp where the mutant allele will have 5 fragments of 249 185 33 24 and 9 bp of size as shown in fig3 size standards used where the 100 bp ladder gibco brl gaithersburg md in an effort to identify a genetic locus responsible for parkinson 39 s disease we performed a genome scan in.