[BCHM 425] - Midterm Exam Guide - Ultimate 18 pages long Study Guide!
5 Oct 2016
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Prepare dna template using spin columns and centriguation. Combine purified plasmid containing cloned gene with seq primers and mail to seq facility. Dna isolation is a process of purification of dna from sample using a comination of physical and chemical methods. The first isolation of dna was done in 1869. 3 basic and 2 optional steps in dna extraction. Cell disttruption or cell lysis to expose dna within. This is commonly achieved by chem and phys methods blending, grinding or sonicating the sample. Remove membrane lipids by adding detergent or surfactants which also serve as cell lysis. Ethanol preiciptation usually by ice-cold ethanol or isopropanol. Since dna insoluble in these alcohols, aggregate tg giving a pellet. Minicolimn purification that relies on the fact that the nucleic acid bind to the solid ohase silicon beads depending on ph and salt content. Targets and amp a single seq from within complex mixture of dna.
