BSC4422 Study guides, Class notes & Summaries

Western blotting a. Proteins from SDS-PAGE are transferred to a nitrocellulose membrane using electrical current a. membrane is blocked so antibody will not bind nonspecifically b. Primary antibody is added and incubated with the membrane c. To detect antibodies that have bound to protein a secondary anti-immunoglobulin antibody coupled to a reporter a. Excess secondary antibody is washed off the membrane b. Substrate is added that precipitates upon reaction c. Reveals visible band where...

Creating DNA Libraries -Collections of cloned DNA fragments from a particular organism contained in plasmid vectors within host bacteria -Screen library to pick out different genes of interest Genomic DNA Libraries -Used to locate fragment with DNA of interest -Chromosomal DNA from tissue of interest is isolated and digested using a restriction enzyme (sticky/blunt ends) -Vector also digested -DNA ligase used to ligate chromosomal DNA fragment inside vector DNA (plasmids) -Recombinant ...