dna分子杂交（DNA molecular hybridization）.docVIP

dna分子杂交（DNA molecular hybridization） Southern blotting: Western blot (Blotting) usually refers to the adsorption of macromolecules or electrophoretic methods by gel electrophoresis separation from the gel onto solid carrier, and then process the probe reaction so as to achieve the specific detection or identification of these macromolecules. Southern first set up DNA to fiber membrane separation from agarose gel electrophoresis in 1975, with specific radioactive isotope labeled DNA (or RNA) fragment hybridization, finally after autoradiography method from X optical film appeared on one or more hybrid zone, thus the detection of specific DNA fragment. Then call this method Southern Blotting (Southern blot); and then in 1977, Alwine more than eight similar methods for detection by agarose gel and polyacrylamide gel electrophoresis (PAGE) separation of specific RNA, this method is also called Northern Blotting (Northern blot) Towbin in 1979; then the method is extended to a specific protein detection and analysis, a Western Blotting (Western blot) PCR: the basic principle of PCR PCR (Polymerase Chain Reaction) is similar to the natural replication process of DNA, and its specificity depends on the oligonucleotide primers complementary to the target sequence. By PCR degeneration, three basic reaction annealing - extension steps: DNA: the denatured template template DNA by heating to 93 DEG C after a certain period of time, the double stranded template DNA amplified by PCR or the formation of double stranded DNA dissociation, to become single, so it and prepare for the next round of primer binding, anti should the template and primer DNA; annealing (renaturation): template DNA by heating denaturation into a single chain, the temperature dropped to 55 degrees Celsius, complementary sequence primer and template DNA strand pairing with the primer extension;: DNA template - primer binding in TaqDNA polymerase under the action of taking dNTP as the raw material, the target se