MIT7_13f08_lab21_Protocol_Spectrophotometric

MIT7_13f08_lab21_Protocol_Spectrophotometric - MIT...

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MIT OpenCourseWare http://ocw.mit.edu 7.13 Experimental Microbial Genetics Fall 2008 For information about citing these materials or our Terms of Use, visit: http://ocw.mit.edu/terms .
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7 . 1 3 S p r i n g 2 0 0 8 P a g e | 1 UV spectrophotometric measurement of DNA concentration and purity DNA itself, and most of the common contaminants found in DNA preps, have absorbances in the region 230nm to 320nm so measurement of the absorbances in this region allows measurement of the DNA concentration and provides information about the contaminant levels. The measurements can be performed on a conventional spectrophotometer, and the most important wavelengths to note are: 230nm: Guanidium salts (used to facilitate DNA binding to silica columns) and phenol (used in phenol/chloroform extractions) absorb strongly at 230nm, therefore high absorbances at this wavelength can be indicative of carry-over of either of these compounds into the sample.
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