design and evaluation PCR primer for type 1 tyrosinemia patient from yasuj

Tyrosinemia is a rare autosomal recessive genetic disease caused by fumarylacetoacetate hydrolase deficiency. 40 different mutation have been recognized related to Tyrosinemia that could be found in all extend of the gene with higher frequency from exon 8 to 14. Because of the size of FAH gene it's impossible to Sequence whole length of the gene by one round of sequencing reaction. Aim of this research is find specific and cost effective primer that could amplify FAH gene in several segments which provide possibility of sequencing of FAH gene. By this approach it's expected to find pathogenic mutation in patient genes. Material and method: first a comprehensive study on pathogenic FAH has been done which revealed the hot spot regions in the gene.  Then for those regions specific primers has been designed by FastPCR software. Selected regions amplified by pcr and sequenced. Finally the sequenced regions analyzed by Vector NTI software. Result: designed primers amplified FAH gene segments specifically and sequencing of amplified segments revealed +709 C>T mutation in exon 10 of FAH gene. Conclusion:  designed primers specifically could be used for diagnosis of carriers and prenatal.