Resources for Genome wide Selection and Mapping of Genes

This project will develop methodology for high-throughout identification of cDNAs from largesegments of genomic DNA cloned into yeast artificial chromosomes (YACs) and from amplified genomicDNA corresponding to a desired micro-dissected chromosome band. We will use cDNA libraries fromhuman spleen, thymus testes, adrenal, thyroid, cerebral cortex, adult brain and fetal brain. We willexplore optimal means of arraying target DNAs in nylon membranes. Hybridization selection will beperformed on the gridded target DNA using as probe the cDNA library pool in the presence of quenchingreagents which block highly abundant or repetitive sequences. Finally, cDNAs hybridizing to a giventarget will be analyzed for repetitive sequences and for genes mapped to the original genomic track. Aset of clones will also be mapped chromosomally and sequenced. The objective is to generatesequencing and mapping information of novel cDNA clones from several genomic locations simultaneous-ly. Hybridization selection is expedient and economizes on cDNA libraries and on quenching reagents.This project should lead to mapped molecular tools for genome wide applications as well as standardreagents and contract services for identification of essentially all human genes.