第六章基因组dna的提取（The sixth chapter is the extraction of genomic DNA）.docVIP

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第六章基因组dna的提取（The sixth chapter is the extraction of genomic DNA）.doc

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第六章基因组dna的提取（The sixth chapter is the extraction of genomic DNA）

第六章基因组dna的提取（The sixth chapter is the extraction of genomic DNA） The sixth chapter is the extraction of genomic DNA The first section is an overview Genomic DNA extraction is usually used to construct genomic libraries, Southern hybrids (including RFLP) and PCR segregating genes. With the longer nature of genomic DNA, it can be separated from small molecules such as organelles or plasmids, DNA. Adding a certain amount of isopropanol or ethanol, high molecular DNA genome or precipitation form fibrous floc floating the available glass rod to remove it, and small molecule DNA can only form granular sediment attached to wall and bottom, so as to achieve the purpose of extraction. In the process of extraction, the chromosome will occur mechanical faults, produce fragments of different size, should be operated under mild conditions so isolating genomic DNA, such as minimizing phenol / chloroform extraction, blending process should be gentle, to get longer DNA. In general, the genomic DNA must be more than 100kb in the construction of genomic libraries. Otherwise, very few fragments with suitable ends are removed on both sides of the enzyme. For RFLP and PCR analysis, the length of DNA can be as short as 50kb, and above this length, RFLP fragments (below 20KB) can be guaranteed after enzyme digestion, and the fragments amplified by PCR (below 2KB) can be guaranteed. Different biological (plant, animal and microorganism) have different extraction methods of genomic DNA; different tissues of different species or the same species due to different cell structure and composition of the separation methods are also different. When extracting the DNA of a particular organization, the corresponding extraction method must be established by referring to the literature and experience to obtain the available DNA macromolecules. In particular, polysaccharides and enzymes in tissues have a strong inhibitory effect on subsequent enzymatic digestion and PCR reaction. Therefore, polysaccha