Strategy to Rapidly Re-Sequence the NF1 Genomic Loci Using Microarrays and Bioinformatics for Molecular Classification of the Disease

摘要

Neurofibromatoses (NF) are disorders caused by mutations in NF1/NF2 genes and characterized by fibromatous tumors on nerves, skin, and bones. A method to rapidly and cheaply re-sequence the NF loci would greatly aid in the molecular classification of the disease by forging links between sequence variations and clinical manifestations. We propose to develop a technique to rapidly re-sequence genomic loci using microarray hybridization and bioinformatics. The NF locus is first amplified using a high processivity polymerase and hybridized on a custom microarray containing all possible 10mer combinations of the four deoxy-ribonucleotides. A virtual profile of a hybridization map of the locus is generated using the available sequence information. This map is then compared to an actual hybridization. If the sequences are identical, then the two images would superimpose. Using the differences in the virtual and real hybridization, we propose to map the mutations in the locus.