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Biotechnology. Analysis of Clonal DNA DNA Cloning Stem Cells Practical Applications of DNA Technology. DNA Cloning Recombinant DNA. Major tool- restriction enzymes; discovered in late 1960’s, cut DNA at restriction site, highly specific, manufactured by bacteria
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Biotechnology Analysis of Clonal DNA DNA Cloning Stem Cells Practical Applications of DNA Technology
DNA Cloning Recombinant DNA • Major tool-restriction enzymes; discovered in late 1960’s, cut DNA at restriction site, highly specific, manufactured by bacteria • Cloning vector-carrier for moving DNA into a cell • http://www.wehi.edu.au/education/wehitv/restriction_enzyme_ecor1/
Procedure for cloning a eukaryotic gene in a bacterial plasmid
How to get a eukaryotic gene to function in a prokaryote • Basic problem: introns!! • Solution: production of cDNA • (artificially produced DNA that is made by using reverse transcriptase on a mRNA)
DNA Libraries • DNA can be cut with restriction enzymes and inserted into plasmids, inserted into bacteria, and stored; whole genome is maintained • cDNA libraries eliminate everything except the genes
PCR-polymerase chain reaction • Rapid amplification of a piece of DNA • Billions of copies in a few hours • Used by CSI, • Jurassic Park? • Prenatal diagnosis
Gel Electrophoresis • Produces DNA “fingerprints” • Separates RFLP’s by using an electrical field • Can be used to identify some genetic diseases • Presence of a particular gene • Used as genetic markers
Other Technologies • Hybridization • Human Genome Project • Physical mapping • Sequencing DNA • Genome Analysis (DNA sequences, gene expression, and gene function)
Stem Cells • Unspecialized cell that can reproduce and differentiate • Types of stem cells Embryonic stem cells from blastula Adult stem cells
Practical Applications of DNA Technology • Disease diagnosis • Gene Therapy • Pharmaceutical products • Forensic uses • Environmental applications • Agricultural applications
Videos and Websites • http://www.pbs.org/wgbh/nova/sciencenow/0302/04.html • http://www.geneticorigins.org/mito/mitoframeset.htm • http://genomics.energy.gov/ • http://www.dnalc.org/ddnalc/resources/cycseq.html • http://www.sumanasinc.com/webcontent/animations/content/pcr.html • http://www.sumanasinc.com/webcontent/animations/content/plasmidcloning.html • http://www.ncbi.nlm.nih.gov/Genbank/index.html
Websites and Videos • http://www.actionbioscience.org/biotech/pecorino2.html • http://www.dnai.org/index.htm • http://people.ibest.uidaho.edu/~etop/vgp/index.html • http://www.dnalc.org/resources/animations/restriction.html • http://www.hhmi.org/biointeractive/dna/animations.html