Manipulating DNA

Manipulating DNA - o multiple cloning site (MCS) - region...

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Manipulating DNA DNA manipulation - uses enzymes (imitates what cells can do) restriction endonuclease - able to cleave DNA at specific places o restriction sites - where nucleases cleave DNA o methylation - stops nucleases from cleaving DNA o Type I - makes simple cuts on both DNA strands o Type II - makes staggered cuts where sequences same on both sides ( dyad symmetry ) ligase - makes phosphodiester bonds to connect hydroxyl/phosphate groups o also joins Okazaki fragments on lagging strands o creates recombinant molecules from fragments created by nucleases vector systems - used to carry recombinant DNA molecule into a cell not required by the cell, but can be selected w/ addition of marker plasmids - small extrachromosomal DNA o must have origin of replication, selectable marker (usually for antibiotic resistance) o markers - used to see which cell took in the new DNA
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Unformatted text preview: o multiple cloning site (MCS) - region in plasmid where DNA is inserted o inactivation of gene signals plasmids acceptance of new DNA phages- viruses that infect bacterial cells o larger than plasmids, can insert more DNA o needs other live cells to replicate o linear DNA (cant infect unless new DNA gets inserted) chimera- totally new genome, nonexistent in nature yeast artificial chromosome (YAC) - able to introduce larger DNA pieces than plasmids DNA library- collection of all DNA fragments representing all an organisms DNA genomic library- simplest type of DNA library o randomly fragmented genome o hydrodynamic shear forces - passes DNA through syringe cDNA libraries- set of all expressed genes o reverse transcriptase- retrovirus that makes DNA from mRNA...
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