toolsofmoleculargenetics-final20141217精编.ppt

The application of tools of Human Molecular Genetics Dan wu Peking University Health Science Center Department of Medical genetic danw@bjmu.edu.cn 分支学科 1．细胞遗传学(cytogenetics) 2．生化遗传学(biochemical genetics) 3．分子遗传学(molecular genetics) 4．群体遗传学(population genetics) 5．药物遗传学(pharmacogenetics) 6．免疫遗传学(immuogenetics) 7．肿瘤遗传学(cancer genetics) 8．毒理遗传学(toxicology genetics) 9. 发育遗传学(developmental genetics) 10. 行为遗传学(behavior genetics) 11. 基因工程(genetic engineering) 12. 优生学(eugenics) 分支学科 1．细胞遗传学(cytogenetics) 2．生化遗传学(biochemical genetics) 3．分子遗传学(molecular genetics) 4．群体遗传学(population genetics) 5．药物遗传学(pharmacogenetics) 6．免疫遗传学(immuogenetics) 7．肿瘤遗传学(cancer genetics) 8．毒理遗传学(toxicology genetics) 9. 发育遗传学(developmental genetics) 10. 行为遗传学(behavior genetics) 11. 基因工程(genetic engineering) 12. 优生学(eugenics) Analysis of individual DNA and  RNA sequences Molecular Cloning Polymerase Chain Reaction (PCR) Restriction Enzymes Example: EcoRⅠ：5’---G^ A A T T C ---3’ 3’---C T T A A ^G ---5’ Sma I : 5’--- CCC ^ GGG ---3’ 3’--- GGG ^ CCC---5’ Vectors Example: plasmid(5000bp), bacteriophage lambda(20kb)，BAC(100-300kb), YAC(1000-2000kb) Molecular Cloning The process of cloning a segment of human DNA into a plasmid cloning vector Bacteriophage Vectors The Polymerase Chain Reaction PCR: Enzymatic amplification of a fragment of DNA located between a pair of primers The Polymerase Chain Reaction The use of molecular cloning and the PCR to isolate large quantities of a particular desired DNA sequence of interest in pure form Construction of Libraries Genomic Libraries Complementary DNA libraries Construction of a “library” of DNA from the human genome in a bacteriophage vector Construction of a cDNA library in a plasmid vector Library Screening Nucleic Acid Probes Probes: a cloned DNA or RNA molecule, labeled