Year: 2015
Type: dissertation
Abstract: Fluorescent dyes that bind DNA have been demonstrated as a useful alternative to radionucleotides for the quantification of DNA and the in vitro measurement of the activity of DNA polymerases and nucleases.However, this approach is generally used in a semi-quantitative way to determine relative rates of reaction.In this report, we demonstrate a method for the simultaneous quantification of DNA in both its single-strand and doublestrand forms using the dye PicoGreen.This approach is used in a steadystate assay of DNA polymerase Klenow fragment exo À , where we determine k cat and K m values for the DNA polymerase that are in excellent agreement with literature values.
Author: Julius Rentergent
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2319603764