rflp限制性内切酶片段长度多态性（Restriction fragment length polymorphism, RFLP）.doc

rflp限制性内切酶片段长度多态性（Restriction fragment length polymorphism, RFLP） First, the basic principle of the The restriction endonuclease (restriction end nuclease) to identify DNA double strand specific DNA sequence and can be cut off. Different restriction endonucleases have their own specific recognition sequences. In the DNA section of the corresponding homologous chromosomes, with a restriction endonuclease digestion, due to different base composition, it will have a different length of the fragment, and then use the corresponding DNA probe labeled with these fragments of good hybridization, showing the map could reflect the genome DNA sequence composed of whether there are differences. This technology is through restriction endonuclease fragment length polymorphism to reveal the similarities and differences between the composition of the DNA sequence, which is called restriction fragment length polymorphism (Restriction Fragment Length Polymorp HISM, abbreviated as RFLP) technology. It can be used to determine whether polymorphism is produced by male or female parent. It can also be used to determine whether the mutation type caused by polymorphism is caused by base mutation or inversion, or by deletion and insertion. The The two basic steps, RFLP Technology The technology mainly includes three steps: the 1, the target DNA to prepare the first genomic DNA extracted and selected the appropriate restriction endonuclease enzyme genomic DNA, with various lengths of DNA fragments were obtained by separation in agarose gel electrophoresis, the fragment length arrangement D NA fragment degeneration after transfer to nitrocellulose membrane or nylon membrane on that blot (Southernb1ot) transfer, And bake at 80 degrees, or with long wave ultraviolet radiation, the DNA fixed on the membrane. The 2, labeled nucleic acid probe will be prepared as a probe of purification of DNA fragments (these DNA fragments can be a fragment of genomic DNA or cDNA, or synthetic oligonucleotides),