第十五章核酸的研究方法.pptxVIP

会计学;1、核酸的分离、提纯和定量测定;DNA的分离 RNA的分离 核酸的定量 ;Isolation of Nucleic Acids;Collection of cells;What’s in Lysis Buffer? Tris buffer to maintain pH of solution so that DNA is stable SDS to dissolve membranes of cell, allowing DNA to be released into solution SDS also denatures proteins, making them more susceptible to protease cleavage;Why add Protease?;Adding salt (5M NaCl);Adding ice cold alcohol;;High MW Genomic DNA Isolation;High MW Genomic DNA Isolation;Isolation of RNA Special Considerations;2、核酸的沉降特性; 通常很难分离出完整DNA分子，但可得到分子量不太大的环状DNA，这类制剂包括： 共价闭环DNA：常呈超螺旋型 开环DNA：双链环状DNA的一条链断裂，分子呈松弛态 线型DNA：环状DNA