DNA定量

作者：Protocol    实验频道来源：本站原创    点击数：    更新时间：2004-7-21

Characterization of dna LEVEL I Materials DNA sample SSC buffer UV spectrophotometer 3 and quartz cuvettes Procedure Dissolve a small quantity of your extracted DNA in 3.0 ml of 0.1X SSC. Turn on and blank a UV spectrophotometer at 220 nm (use 0.1X SSC as the blank). Determine the absorbance of your sample DNA at 230 nm. Change the wavelength to 230 nm, reblank the spectrophotometer and measure the absorbance of the sample at 230 nm. Increment the wavelength by 10 nm and repeat blanking and measuring the absorbance until readings are taken through 300 nm. Compute the absorbance ratio 260 nm to 280 nm. Pure DNA (without protein or RNA) will have a 260:280 absorbance ratio of 1.85. RNA will have a 260:280 ratio of 2.0. Plot the absorbance spectrum of your sample and indicate the 260:280 ratio, as well as the amount of protein contamination on the graph. Wavelength Absorbance 220 230 240 250 260 270 280 290 300 处理 SSI 文件时出错