北京大学分子生物学课件molecular-biology.pptVIP

祝同学们新年快乐！考试顺利！ * * * * * * * * * * Positional cloning is powerful, but it has also been extremely tedious. When the approach was first proposed in the early 1980s9, a researcher wishing to perform positional cloning had to generate genetic markers to trace inheritance; perform chromosomal walking to obtain genomic DNA covering the region; and analyse a region of around 1 Mb by either direct sequencing or indirect gene identification methods. The first two barriers were eliminated with the development in the mid-1990s of comprehensive genetic and physical maps of the human chromosomes, under the auspices of the Human Genome Project. The remaining barrier, however, has continued to be formidable. * Produce finished sequence from the clones spanning the physical map, close gaps, develop new technologies for difficult areas Comparison to other genomes to identify potential exons – sequence is not enough Expression arrays Already, projects are underway to sequence the genomes of the mouse, rat, zebrafish and the pufferfishes. What to next should be based on phylogeny, relevance to understanding human biology and medicine, economic importance – agriculture rice, cow, genome characteristics including size, chromosome conservation, species diversity The human draft genome sequence has already allowed the identification of more than 1.4 million SNPs, comprising a substantial proportion of all common human variation. This program should be extended to obtain a nearly complete catalogue of common variants and to identify the common ancestral haplotypes present in the population. The scientific program outlined above focuses on how the genome sequence can be mined for biological information. In addition, the sequence will serve as a foundation for a broad range of functional genomic tools to help biologists to probe function in a more systematic manner. Fragment DNA and ligate adaptors Comparison WGS 454 Solexa Cloning Yes No No Chemistry Sanger pyrosequencing reversible