This is a proposal for funds to acquire a multi-task system for automated analysis of mutations and polymorphisms and for drastically expanded capacity of automated DNA sequencing. The funds are intended to upgrade an existing DNA sequencer as well as to purchase a second one. The funds will also be used to purchase a robotic lab station that automates cycle sequencing reactions and software that analyzes complex electrophoretic DNA patterns. Automated analysis and quantitation of complex DNA electrophoretic patterns at nucleotide resolution is necessary for microsatellite, ARMS PCR and SSCP analyses of polymorphic loci. Expanded DNA sequencing capacity is vital to current studies of DNA sequence analysis including transcriptional regulatory regions of cell-specific genes, cDNA clones for transcription factors and their interacting proteins and autoantibodies, verifying site-directed mutations to study structure-function relationships in proteins and DNA transcriptional control elements, the nucleotide basis of genetic polymorphisms, chromosomal breakpoints implicated in oncogene activation, analysis of extended loci of multigene families and sequence verification of extended transgenes for introduction into mice.
