Geneid--A New Gene Mapping and Finding System

This proposal describes a set of reagents which will make gene mapping to chromosomes and chromosomas well as gene finding exercises, accessible to any molecular biologist or geneticist familiar withtechniques. These reagents, called the Gene. ID system, are a set of "dot-blots". The "Chromosome.IDcorresponding to each chromosome spotted in a dot-blot format and the "Band.ID" blot contains DNA coeach chromosome band also spotted in a dot blot format. These chromosome-specific and band-specificprepared by dissecting and pooling 10 copies of each chromosome, or each chromosome band, followed bamplification using a universal degenerate primer. The primer is 21 based long and contains 10 nucleDNA, so that at low annealing and extension temperatures, virtually any DNA is amplifiable. These fimap any cloned DNA to a chromosome or a chromosome band with standard filter hybridization techniquefeasibility demonstration of the Chromosome.ID and Band.ID blots will consist of preparation of seveChromosome.ID that contain chromosome- specific DNA prepared from mouse chromosomes 11, along with cchromosomes 2 and 9. These blots will be hybridized with a series of single copy gene probes known tchromosome 11. Conditions for mapping complex probes such as cosmids, YACs, or lambda clones will alinvestigated, thus increasing the spectrum of potential users. Finally, the feasibility of physicallsequences from the chromosome-specific DNAs prior to application to the Gene. ID blots will also bePhase I. Removal of the repetitive sequences ultimately will make the blots more "user friendly" becwill allow the users to hybridize their probes without prior suppression of repetitive sequences usi