| 摘要 |
PURPOSE: Provided is a method for separating mitochondrial DAN(mtDNA) from a small amount of animal tissues, thereby separating pure mtDNA from a large amount of animal tissues or cells without using chemicals, such as phenol and the like. CONSTITUTION: The separation method of mtDNA from a large amount of animal tissues comprises the steps of: adding a first buffer to a sample tissue in a mortar; transferring the first buffer to a homogenizing tube with a gauze then homogenizing it; dividing the supernatant into two test tube then followed by centrifuging them and transferring supernatants to another test tubes each; removing the supernatant then adding a second buffer to the each test tube with mixing; transferring the resultants to another test tubes each and centrifuging them; leaving a small amount of the supernatant each with stirring; adding a third buffer thereto with stirring then leaving them on ice; adding a forth buffer to the resultants with stirring then leaving them on ice; centrifuging them and transferring supernatants to another test tubes each; adding a fifth and a sixth buffer to the resultants with stirring then centrifuging them to remove supernatants; adding the fifth buffer to the resultants with stirring then removing supernatants; adding seventh buffer to the resultants with stirring then removing supernatants; drying the resultants with a vacuum pump; and adding an eighth buffer the resultant then followed by centrifuging and transferring supernatants to new clean test tubes each.
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