Solid-phase plate-reader quantification of specific PCR products by measurement of band-specific ethidium bromide fluorescence.
Real-time PCR is widely employed to quantify PCR products across a range of applications. However, accurate real-time PCR is not always technically feasible, and alternative methods for PCR product quantification can be expensive and time consuming to validate. We have developed an inexpensive, rapid, and immediately accessible protocol to quantify PCR products, by measuring ethidium bromide fluorescence of PCR products excised from agarose gels. This protocol has relevance to a broad range of methods in molecular biology where quantification of PCR products is necessary.
Centre for Cellular and Molecular Physiology, University of Oxford, Oxford OX3 7BN, UK.
Michael T
MT
McCarthy
Centre for Cellular and Molecular Physiology, University of Oxford, Oxford OX3 7BN, UK. Electronic address: chris.ocallaghan@ndm.ox.ac.uk.
Christopher A
CA
O'Callaghan
D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004996:Ethidium / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D013050:Spectrometry, Fluorescence
D004247:DNA; D004996:Ethidium
Ethidium bromide; Fluorescence; PCR; Quantification