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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 double-strand forms using the dye PicoGreen. This approach is used in a steady-state assay of DNA polymerase Klenow fragment exo???, where we determine kcat and Km values for the DNA polymerase that are in excellent agreement with literature values.
Original language | English |
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Pages (from-to) | 2042-2050 |
Number of pages | 9 |
Journal | FEBS Journal |
Volume | 281 |
Issue number | 8 |
DOIs | |
Publication status | Published - Apr 2014 |
Keywords
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Dive into the research topics of 'A quantitative fluorescence-based steady-state assay of DNA polymerase'. Together they form a unique fingerprint.Projects
- 1 Finished
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Linking experiment to theory: Quantum entanglement during enzyme catalysis - Dr S Hay fellowship
Hay, S. (PI)
1/09/10 → 31/08/15
Project: Research