Manipulating DNA

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

Info iconThis preview shows page 1. Sign up to view the full content.

View Full Document Right Arrow Icon
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
Background image of page 1
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: o multiple cloning site (MCS) - region in plasmid where DNA is inserted o inactivation of gene signals plasmid’s 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 (can’t 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 organism’s 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...
View Full Document

{[ snackBarMessage ]}

Ask a homework question - tutors are online