Sisällysluettelo:
“…Chapter 5 DNA Quantification Paul
A. Heaton and Jacquie T. Keer 5.1 Introduction 83 5.
2 Measurement of DNA Concentration Using Ultraviolet Spectroscopy 84 5.
2.1 Determining the Extinction Coefficient e 84 5.
2.
2 Practical Aspects of Measuring DNA Concentrations by UV Spectroscopy 85 5.
2.
2.1 Calibration of the Spectrophotometer 85 5.
2.
2.
2 Cuvettes 85 5.
2.
2.3 Sample Preparation 86 5.
2.
2.4 Reference Blank 86 5.
2.
2.5 Sample Dilution 86 5.
2.
2.6 Light Source 86 5.
2.
2.7 Presence of Contaminants 87 5.3 Determination of DNA Concentration by Fluorescence Spectroscopy 88 5.3.1 Preparation of
a Calibration Graph 88 5.3.
2 Practical Aspects of Measuring DNA Concentrations by Fluorescence Spectroscopy 89 5.3.
2.1 Sample Preparation 89 5.3.
2.
2 Reference Blank 90 5.3.
2.3 DNA Standard 90 5.3.
2.4 Selecting the Dye 90 5.3.
2.5 Dye Concentration 90 5.3.
2.6 Microtitre Plates 90 5.3.
2.7 Measurement Conditions 90 5.3.3 Fluorescent Dyes 91 5.3.3.1 Ethidium Bromide 91 5.3.3.
2 PicoGreens 92 5.3.3.3 SYBR Dyes 92 5.3.3.4 Hoechst 33258 92 5.4 Quantification Using the Polymerase Chain Reaction 93 5.5 Enzymatic Quantification of DNA 93 5.6 Primary Methods of DNA Quantification 94 5.6.1 Gravimetric Analysis 95 5.6.
2 Isotope Dilution Mass Spectrometry for Oligonucleotide Quantification 95 xii Contents 5.7 DNA Quantification by Constituent Phosphorus Determination 97 5.8 Comparability of DNA Measurement Methods 98 5.9 Summary 99 References…”
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