Objective The objectives are: to construct cosmid libraries from flow sorted human chromosomes 17 and 11 (at least 5 genome equivalents each); to pick individual clones into microtitre plate wells for storage and reference and generate at least two duplicate microtitre plates of the entire clone set for handling in the reference library system; to produce high density membranes on request from chromosome specific libraries (approximately 10 000 clones per membrane) on which the clones from microtitre plate wells are grown as colonies, lysed in situ, and converted into DNA for hybridization. Each chromosome specific library set is represented by one or two membranes bearing 4 to 5 genomic equivalents of cloned DNA, giving a high (80 to 90%) probability to detect clones from any single chromosomal locus used as a probe. This will include the production of high density membranes for the screening requests of chromosomes 11 and 17 as well as for the new requests of the already existing libraries for chromosomes X and 21; to distribute up to 50 (taking in account all four chromosomes), individual chromosome library set requests per year (a total of about 90 individual high density membranes per year have to be produced, allowing for some imperfect membranes and duplications) to the research groups interested in screening these libraries; to serve as a reference source of cosmid clones for human chromosome specific libraries (chromosomes 17, 11, X and 21) with a capacity to provide up to 2000 clones per year; to organize a central database which will integrate information on all probes used to screen the libraries and assign it to individual clones. These clones (and the probe details) would then be available for distribution either generally (if detected with published probes), or locally to the researchers who detected them (if detected with unpublished probes). The integrated mapping of the target genomes by integration of data obtained through hybridization using unique and complex robes has been a long term strategy of the Genome Analysis Laboratory (GAL) at the Imperial Cancer Research Fund (ICRF). Cosmid libraries have been constructed from flow sorted human chromosomes X and human chromosomes 21 each of which contained over 30 genomic equivalents. These had been picked into microtire plates and duplicated. Libraries had been characterized both by analysing random clones and screening with genetically mapped probes. High density membranes have been prepared from microtitre plates such that 2 membranes or 1 membrane bear 4 to 5 genome equivalents of chromosomes X and 21, respectively. Such membranes have been distributed to more than 40 different laboratories (each chromosome) and over 100 single copy probes have been hybridized (the membranes for each chromosomes detecting in more than 90% of cases, 1 to 10 positive cosmids per probe). The total number of positive clones detected up to a year ago on both libraries exceeded 4000. Additional cosmid libraries are being constructed for the human chromosomes 11 and human chromosomes 17 and the distribution of clones and membranes for 4 chromosomes (11, 17, 21, X). Cosmid libraries for chromosomes 11 and 17, as well as additional chromosomes 6, chromosomes 13, chromosomes 18 and chromosomes 22 have been running for different time periods as reference libraries and 514 high density membrane sets have been sent to different laboratories in 10 European countries as well as a number of laboratories on other continents. A total of 1266 probes have detected over 10 000 positive clones that have been sent to users as requested in 1508 individual requests. For about 80% of these deliverables the beneficiaries have been the European laboratories. Several contigs in the genetically important regions have been identified. Fields of science natural sciencescomputer and information sciencesdatabasesnatural sciencesbiological sciencesgeneticsDNAmedical and health sciencesclinical medicineoncologynatural sciencesbiological sciencesgeneticschromosomesnatural sciencesbiological sciencesgeneticsgenomes Programme(s) FP2-HUMGEN C - Specific research programme (EEC) in the field of health: human genome analysis, 1990-1992 Topic(s) Data not available Call for proposal Data not available Funding Scheme CSC - Cost-sharing contracts Coordinator Imperial Cancer Research Fund (ICRF) EU contribution No data Address 44 Lincoln's Inn Fields WC2A 3PX London United Kingdom See on map Total cost No data
