G Protein-Coupled Receptors (eBook, PDF)
Essential Methods
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Essential Methods
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G-Protein Coupled Receptors (GPCRs) are not only the largest protein family in the human genome but are also the single biggest target for therapeutic agents. Research into GPCRs is therefore growing at a fast pace and the range of techniques that can be applied to GPCRs is vast and continues to grow. This book provides an invaluable bench-side guide into the best and most up-to-date techniques for current and future research on GPCRs. With contributions from leading international authorities, this book equips readers with clear and detailed protocols for both well-known and up-and-coming…mehr
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G-Protein Coupled Receptors (GPCRs) are not only the largest protein family in the human genome but are also the single biggest target for therapeutic agents. Research into GPCRs is therefore growing at a fast pace and the range of techniques that can be applied to GPCRs is vast and continues to grow. This book provides an invaluable bench-side guide into the best and most up-to-date techniques for current and future research on GPCRs. With contributions from leading international authorities, this book equips readers with clear and detailed protocols for both well-known and up-and-coming techniques along with hints and tips for success. All the methods have been tried and tested by leading international research labs and are presented in easy-to-follow stages along with a useful overview of each technique. This book is an essential resource for all researchers in molecular biology, biochemistry, pharmacology and for graduate students.
Produktdetails
- Produktdetails
- Verlag: John Wiley & Sons
- Seitenzahl: 310
- Erscheinungstermin: 28. November 2009
- Englisch
- ISBN-13: 9780470749227
- Artikelnr.: 37299289
- Verlag: John Wiley & Sons
- Seitenzahl: 310
- Erscheinungstermin: 28. November 2009
- Englisch
- ISBN-13: 9780470749227
- Artikelnr.: 37299289
Dr David R. Poyner, University of Aston, Birmingham, UK; School of Life and Health Sciences, Aston University, Birmingham, UK. Professor?Mark Wheatley, University of Birmingham, UK.
Preface. Contributors. 1 Measurement of Ligand-G-Protein-coupled ReceptorInteractions (Katie Leach, Celine Valant, Patrick M. Sextonand Arthur Christopoulos). 1.1 Introduction. 1.2 Methods and Approaches. References. 2 Second Messenger Assays for G-Protein-coupled Receptors:cAMP, Ca¯2+, Inositol Phosphates¯, ERK1/2(Karen J. Gregory, Patrick M. Sexton, Arthur Christopoulosand Caroline A. Hick). 2.1 Introduction. 2.2 Methods and Approaches. 2.3 Troubleshooting. References. 3 Use of the [¯35S]GTPgammaS Binding Assayto Determine Ligand Efficacy at G-protein-coupled Receptors(Elodie Kara and Philip G. Strange). 3.1 Introduction. 3.2 Methods and Approaches. 3.3 Troubleshooting. Acknowledgements. References. 4 Quantitative Imaging of Receptor Trafficking (AndyR. James, Takeo Awaji, F. Anne Stephenson and Nicholas A.Hartell). 4.1 Introduction. 4.2 Methods and Approaches. 4.3 Troubleshooting. References. 5 Production of Recombinant G-Protein-coupled Receptor inYeast for Structural and Functional Analysis (Richard A.J.Darby, Mohammed Jamshad, Ljuban Grgic, William J. Holmes and RoslynM. Bill). 5.1 Introduction. 5.2 Methods and Approaches. 5.3 Troubleshooting. References. 6 Monitoring GPCR-Protein Complexes UsingBioluminescence Resonance Energy Transfer (Werner C. Jaeger,Kevin D.G. Pfleger and Karin A. Eidne). 6.1 Introduction. 6.2 Methods and Approaches. 6.3 Troubleshooting. References. 7 Using Intramolecular Fluorescence Resonance Energy Transferto Study Receptor Conformation (Cornelius Krasel and CarstenHoffmann). 7.1 Introduction. 7.2 Methods and Approaches. 7.3 Troubleshooting. References. 8 A Disulfide Cross-linking Strategy Useful for StudyingLigand-induced Structural Changes in GPCRs (Jian Hua Li,Stuart D.C. Ward, Sung-Jun Han, Fadi F. Hamdan and JürgenWess). 8.1 Introduction. 8.2 Methods and Approaches. 8.3 General Considerations, Caveats and Troubleshooting. Acknowledgements. References. 9 Use of Fluorescence Correlation Spectroscopy to Study theDiffusion of G-protein-coupled Receptors (Stephen J.Briddon, Jonathan A. Hern and Stephen J. Hill). 9.1 Introduction. 9.2 Methods and Approaches. 9.3 Troubleshooting. References. 10 Identification and Proteomic Analysis of GPCRPhosphorylation (Kok Choi Kong, Sharad C. Mistry and AndrewB. Tobin). 10.1 Introduction. 10.2 Methods. Acknowledgements. References. 11 Measurement and Visualization of G-Protein-coupledReceptor Trafficking by Enzyme-linked Immunosorbent Assay andImmunofluorescence (Stuart J. Mundell, Shaista P. Nisar andEamonn Kelly). 11.1 Introduction. 11.2 Methods and Approaches. 11.3 Troubleshooting. References. 12 Substituted Cysteine Accessibility Method (SCAM)(George Liapakis and Jonathan A. Javitch). 12.1 Introduction. 12.2 Methods and Approaches. 12.3 Troubleshooting. References. 13 Homology Modelling of G-Protein-coupled Receptors(John Simms). 13.1 Introduction. 13.2 Methods and Approaches. 13.3 Troubleshooting. 13.4 Automated Methods for Generating Models of GPCRs. References. Appendix: Site-directed Mutagenesis and Chimeras (AlexConner, Mark Wheatley and David Poyner). A.1 Introduction. A.2 Why Mutagenesis? A.3 Troubleshooting. A.4 Conclusion. References. Index.
Preface. Contributors. 1 Measurement of Ligand-G-Protein-coupled Receptor
Interactions (Katie Leach, Celine Valant, Patrick M. Sexton and Arthur
Christopoulos). 1.1 Introduction. 1.2 Methods and Approaches. References. 2
Second Messenger Assays for G-Protein-coupled Receptors: cAMP, Ca2+,
Inositol Phosphates, ERK1/2 (Karen J. Gregory, Patrick M. Sexton, Arthur
Christopoulos and Caroline A. Hick). 2.1 Introduction. 2.2 Methods and
Approaches. 2.3 Troubleshooting. References. 3 Use of the [35S]GTP³S
Binding Assay to Determine Ligand Efficacy at G-protein-coupled Receptors
(Elodie Kara and Philip G. Strange). 3.1 Introduction. 3.2 Methods and
Approaches. 3.3 Troubleshooting. Acknowledgements. References. 4
Quantitative Imaging of Receptor Trafficking (Andy R. James, Takeo Awaji,
F. Anne Stephenson and Nicholas A. Hartell). 4.1 Introduction. 4.2 Methods
and Approaches. 4.3 Troubleshooting. References. 5 Production of
Recombinant G-Protein-coupled Receptor in Yeast for Structural and
Functional Analysis (Richard A.J. Darby, Mohammed Jamshad, Ljuban Grgic,
William J. Holmes and Roslyn M. Bill). 5.1 Introduction. 5.2 Methods and
Approaches. 5.3 Troubleshooting. References. 6 Monitoring GPCR-Protein
Complexes Using Bioluminescence Resonance Energy Transfer (Werner C.
Jaeger, Kevin D.G. Pfleger and Karin A. Eidne). 6.1 Introduction. 6.2
Methods and Approaches. 6.3 Troubleshooting. References. 7 Using
Intramolecular Fluorescence Resonance Energy Transfer to Study Receptor
Conformation (Cornelius Krasel and Carsten Hoffmann). 7.1 Introduction. 7.2
Methods and Approaches. 7.3 Troubleshooting. References. 8 A Disulfide
Cross-linking Strategy Useful for Studying Ligand-induced Structural
Changes in GPCRs (Jian Hua Li, Stuart D.C. Ward, Sung-Jun Han, Fadi F.
Hamdan and Jürgen Wess). 8.1 Introduction. 8.2 Methods and Approaches. 8.3
General Considerations, Caveats and Troubleshooting. Acknowledgements.
References. 9 Use of Fluorescence Correlation Spectroscopy to Study the
Diffusion of G-protein-coupled Receptors (Stephen J. Briddon, Jonathan A.
Hern and Stephen J. Hill). 9.1 Introduction. 9.2 Methods and Approaches.
9.3 Troubleshooting. References. 10 Identification and Proteomic Analysis
of GPCR Phosphorylation (Kok Choi Kong, Sharad C. Mistry and Andrew B.
Tobin). 10.1 Introduction. 10.2 Methods. Acknowledgements. References. 11
Measurement and Visualization of G-Protein-coupled Receptor Trafficking by
Enzyme-linked Immunosorbent Assay and Immunofluorescence (Stuart J.
Mundell, Shaista P. Nisar and Eamonn Kelly). 11.1 Introduction. 11.2
Methods and Approaches. 11.3 Troubleshooting. References. 12 Substituted
Cysteine Accessibility Method (SCAM) (George Liapakis and Jonathan A.
Javitch). 12.1 Introduction. 12.2 Methods and Approaches. 12.3
Troubleshooting. References. 13 Homology Modelling of G-Protein-coupled
Receptors (John Simms). 13.1 Introduction. 13.2 Methods and Approaches.
13.3 Troubleshooting. 13.4 Automated Methods for Generating Models of
GPCRs. References. Appendix: Site-directed Mutagenesis and Chimeras (Alex
Conner, Mark Wheatley and David Poyner). A.1 Introduction. A.2 Why
Mutagenesis? A.3 Troubleshooting. A.4 Conclusion. References. Index.
Interactions (Katie Leach, Celine Valant, Patrick M. Sexton and Arthur
Christopoulos). 1.1 Introduction. 1.2 Methods and Approaches. References. 2
Second Messenger Assays for G-Protein-coupled Receptors: cAMP, Ca2+,
Inositol Phosphates, ERK1/2 (Karen J. Gregory, Patrick M. Sexton, Arthur
Christopoulos and Caroline A. Hick). 2.1 Introduction. 2.2 Methods and
Approaches. 2.3 Troubleshooting. References. 3 Use of the [35S]GTP³S
Binding Assay to Determine Ligand Efficacy at G-protein-coupled Receptors
(Elodie Kara and Philip G. Strange). 3.1 Introduction. 3.2 Methods and
Approaches. 3.3 Troubleshooting. Acknowledgements. References. 4
Quantitative Imaging of Receptor Trafficking (Andy R. James, Takeo Awaji,
F. Anne Stephenson and Nicholas A. Hartell). 4.1 Introduction. 4.2 Methods
and Approaches. 4.3 Troubleshooting. References. 5 Production of
Recombinant G-Protein-coupled Receptor in Yeast for Structural and
Functional Analysis (Richard A.J. Darby, Mohammed Jamshad, Ljuban Grgic,
William J. Holmes and Roslyn M. Bill). 5.1 Introduction. 5.2 Methods and
Approaches. 5.3 Troubleshooting. References. 6 Monitoring GPCR-Protein
Complexes Using Bioluminescence Resonance Energy Transfer (Werner C.
Jaeger, Kevin D.G. Pfleger and Karin A. Eidne). 6.1 Introduction. 6.2
Methods and Approaches. 6.3 Troubleshooting. References. 7 Using
Intramolecular Fluorescence Resonance Energy Transfer to Study Receptor
Conformation (Cornelius Krasel and Carsten Hoffmann). 7.1 Introduction. 7.2
Methods and Approaches. 7.3 Troubleshooting. References. 8 A Disulfide
Cross-linking Strategy Useful for Studying Ligand-induced Structural
Changes in GPCRs (Jian Hua Li, Stuart D.C. Ward, Sung-Jun Han, Fadi F.
Hamdan and Jürgen Wess). 8.1 Introduction. 8.2 Methods and Approaches. 8.3
General Considerations, Caveats and Troubleshooting. Acknowledgements.
References. 9 Use of Fluorescence Correlation Spectroscopy to Study the
Diffusion of G-protein-coupled Receptors (Stephen J. Briddon, Jonathan A.
Hern and Stephen J. Hill). 9.1 Introduction. 9.2 Methods and Approaches.
9.3 Troubleshooting. References. 10 Identification and Proteomic Analysis
of GPCR Phosphorylation (Kok Choi Kong, Sharad C. Mistry and Andrew B.
Tobin). 10.1 Introduction. 10.2 Methods. Acknowledgements. References. 11
Measurement and Visualization of G-Protein-coupled Receptor Trafficking by
Enzyme-linked Immunosorbent Assay and Immunofluorescence (Stuart J.
Mundell, Shaista P. Nisar and Eamonn Kelly). 11.1 Introduction. 11.2
Methods and Approaches. 11.3 Troubleshooting. References. 12 Substituted
Cysteine Accessibility Method (SCAM) (George Liapakis and Jonathan A.
Javitch). 12.1 Introduction. 12.2 Methods and Approaches. 12.3
Troubleshooting. References. 13 Homology Modelling of G-Protein-coupled
Receptors (John Simms). 13.1 Introduction. 13.2 Methods and Approaches.
13.3 Troubleshooting. 13.4 Automated Methods for Generating Models of
GPCRs. References. Appendix: Site-directed Mutagenesis and Chimeras (Alex
Conner, Mark Wheatley and David Poyner). A.1 Introduction. A.2 Why
Mutagenesis? A.3 Troubleshooting. A.4 Conclusion. References. Index.
Preface. Contributors. 1 Measurement of Ligand-G-Protein-coupled ReceptorInteractions (Katie Leach, Celine Valant, Patrick M. Sextonand Arthur Christopoulos). 1.1 Introduction. 1.2 Methods and Approaches. References. 2 Second Messenger Assays for G-Protein-coupled Receptors:cAMP, Ca¯2+, Inositol Phosphates¯, ERK1/2(Karen J. Gregory, Patrick M. Sexton, Arthur Christopoulosand Caroline A. Hick). 2.1 Introduction. 2.2 Methods and Approaches. 2.3 Troubleshooting. References. 3 Use of the [¯35S]GTPgammaS Binding Assayto Determine Ligand Efficacy at G-protein-coupled Receptors(Elodie Kara and Philip G. Strange). 3.1 Introduction. 3.2 Methods and Approaches. 3.3 Troubleshooting. Acknowledgements. References. 4 Quantitative Imaging of Receptor Trafficking (AndyR. James, Takeo Awaji, F. Anne Stephenson and Nicholas A.Hartell). 4.1 Introduction. 4.2 Methods and Approaches. 4.3 Troubleshooting. References. 5 Production of Recombinant G-Protein-coupled Receptor inYeast for Structural and Functional Analysis (Richard A.J.Darby, Mohammed Jamshad, Ljuban Grgic, William J. Holmes and RoslynM. Bill). 5.1 Introduction. 5.2 Methods and Approaches. 5.3 Troubleshooting. References. 6 Monitoring GPCR-Protein Complexes UsingBioluminescence Resonance Energy Transfer (Werner C. Jaeger,Kevin D.G. Pfleger and Karin A. Eidne). 6.1 Introduction. 6.2 Methods and Approaches. 6.3 Troubleshooting. References. 7 Using Intramolecular Fluorescence Resonance Energy Transferto Study Receptor Conformation (Cornelius Krasel and CarstenHoffmann). 7.1 Introduction. 7.2 Methods and Approaches. 7.3 Troubleshooting. References. 8 A Disulfide Cross-linking Strategy Useful for StudyingLigand-induced Structural Changes in GPCRs (Jian Hua Li,Stuart D.C. Ward, Sung-Jun Han, Fadi F. Hamdan and JürgenWess). 8.1 Introduction. 8.2 Methods and Approaches. 8.3 General Considerations, Caveats and Troubleshooting. Acknowledgements. References. 9 Use of Fluorescence Correlation Spectroscopy to Study theDiffusion of G-protein-coupled Receptors (Stephen J.Briddon, Jonathan A. Hern and Stephen J. Hill). 9.1 Introduction. 9.2 Methods and Approaches. 9.3 Troubleshooting. References. 10 Identification and Proteomic Analysis of GPCRPhosphorylation (Kok Choi Kong, Sharad C. Mistry and AndrewB. Tobin). 10.1 Introduction. 10.2 Methods. Acknowledgements. References. 11 Measurement and Visualization of G-Protein-coupledReceptor Trafficking by Enzyme-linked Immunosorbent Assay andImmunofluorescence (Stuart J. Mundell, Shaista P. Nisar andEamonn Kelly). 11.1 Introduction. 11.2 Methods and Approaches. 11.3 Troubleshooting. References. 12 Substituted Cysteine Accessibility Method (SCAM)(George Liapakis and Jonathan A. Javitch). 12.1 Introduction. 12.2 Methods and Approaches. 12.3 Troubleshooting. References. 13 Homology Modelling of G-Protein-coupled Receptors(John Simms). 13.1 Introduction. 13.2 Methods and Approaches. 13.3 Troubleshooting. 13.4 Automated Methods for Generating Models of GPCRs. References. Appendix: Site-directed Mutagenesis and Chimeras (AlexConner, Mark Wheatley and David Poyner). A.1 Introduction. A.2 Why Mutagenesis? A.3 Troubleshooting. A.4 Conclusion. References. Index.
Preface. Contributors. 1 Measurement of Ligand-G-Protein-coupled Receptor
Interactions (Katie Leach, Celine Valant, Patrick M. Sexton and Arthur
Christopoulos). 1.1 Introduction. 1.2 Methods and Approaches. References. 2
Second Messenger Assays for G-Protein-coupled Receptors: cAMP, Ca2+,
Inositol Phosphates, ERK1/2 (Karen J. Gregory, Patrick M. Sexton, Arthur
Christopoulos and Caroline A. Hick). 2.1 Introduction. 2.2 Methods and
Approaches. 2.3 Troubleshooting. References. 3 Use of the [35S]GTP³S
Binding Assay to Determine Ligand Efficacy at G-protein-coupled Receptors
(Elodie Kara and Philip G. Strange). 3.1 Introduction. 3.2 Methods and
Approaches. 3.3 Troubleshooting. Acknowledgements. References. 4
Quantitative Imaging of Receptor Trafficking (Andy R. James, Takeo Awaji,
F. Anne Stephenson and Nicholas A. Hartell). 4.1 Introduction. 4.2 Methods
and Approaches. 4.3 Troubleshooting. References. 5 Production of
Recombinant G-Protein-coupled Receptor in Yeast for Structural and
Functional Analysis (Richard A.J. Darby, Mohammed Jamshad, Ljuban Grgic,
William J. Holmes and Roslyn M. Bill). 5.1 Introduction. 5.2 Methods and
Approaches. 5.3 Troubleshooting. References. 6 Monitoring GPCR-Protein
Complexes Using Bioluminescence Resonance Energy Transfer (Werner C.
Jaeger, Kevin D.G. Pfleger and Karin A. Eidne). 6.1 Introduction. 6.2
Methods and Approaches. 6.3 Troubleshooting. References. 7 Using
Intramolecular Fluorescence Resonance Energy Transfer to Study Receptor
Conformation (Cornelius Krasel and Carsten Hoffmann). 7.1 Introduction. 7.2
Methods and Approaches. 7.3 Troubleshooting. References. 8 A Disulfide
Cross-linking Strategy Useful for Studying Ligand-induced Structural
Changes in GPCRs (Jian Hua Li, Stuart D.C. Ward, Sung-Jun Han, Fadi F.
Hamdan and Jürgen Wess). 8.1 Introduction. 8.2 Methods and Approaches. 8.3
General Considerations, Caveats and Troubleshooting. Acknowledgements.
References. 9 Use of Fluorescence Correlation Spectroscopy to Study the
Diffusion of G-protein-coupled Receptors (Stephen J. Briddon, Jonathan A.
Hern and Stephen J. Hill). 9.1 Introduction. 9.2 Methods and Approaches.
9.3 Troubleshooting. References. 10 Identification and Proteomic Analysis
of GPCR Phosphorylation (Kok Choi Kong, Sharad C. Mistry and Andrew B.
Tobin). 10.1 Introduction. 10.2 Methods. Acknowledgements. References. 11
Measurement and Visualization of G-Protein-coupled Receptor Trafficking by
Enzyme-linked Immunosorbent Assay and Immunofluorescence (Stuart J.
Mundell, Shaista P. Nisar and Eamonn Kelly). 11.1 Introduction. 11.2
Methods and Approaches. 11.3 Troubleshooting. References. 12 Substituted
Cysteine Accessibility Method (SCAM) (George Liapakis and Jonathan A.
Javitch). 12.1 Introduction. 12.2 Methods and Approaches. 12.3
Troubleshooting. References. 13 Homology Modelling of G-Protein-coupled
Receptors (John Simms). 13.1 Introduction. 13.2 Methods and Approaches.
13.3 Troubleshooting. 13.4 Automated Methods for Generating Models of
GPCRs. References. Appendix: Site-directed Mutagenesis and Chimeras (Alex
Conner, Mark Wheatley and David Poyner). A.1 Introduction. A.2 Why
Mutagenesis? A.3 Troubleshooting. A.4 Conclusion. References. Index.
Interactions (Katie Leach, Celine Valant, Patrick M. Sexton and Arthur
Christopoulos). 1.1 Introduction. 1.2 Methods and Approaches. References. 2
Second Messenger Assays for G-Protein-coupled Receptors: cAMP, Ca2+,
Inositol Phosphates, ERK1/2 (Karen J. Gregory, Patrick M. Sexton, Arthur
Christopoulos and Caroline A. Hick). 2.1 Introduction. 2.2 Methods and
Approaches. 2.3 Troubleshooting. References. 3 Use of the [35S]GTP³S
Binding Assay to Determine Ligand Efficacy at G-protein-coupled Receptors
(Elodie Kara and Philip G. Strange). 3.1 Introduction. 3.2 Methods and
Approaches. 3.3 Troubleshooting. Acknowledgements. References. 4
Quantitative Imaging of Receptor Trafficking (Andy R. James, Takeo Awaji,
F. Anne Stephenson and Nicholas A. Hartell). 4.1 Introduction. 4.2 Methods
and Approaches. 4.3 Troubleshooting. References. 5 Production of
Recombinant G-Protein-coupled Receptor in Yeast for Structural and
Functional Analysis (Richard A.J. Darby, Mohammed Jamshad, Ljuban Grgic,
William J. Holmes and Roslyn M. Bill). 5.1 Introduction. 5.2 Methods and
Approaches. 5.3 Troubleshooting. References. 6 Monitoring GPCR-Protein
Complexes Using Bioluminescence Resonance Energy Transfer (Werner C.
Jaeger, Kevin D.G. Pfleger and Karin A. Eidne). 6.1 Introduction. 6.2
Methods and Approaches. 6.3 Troubleshooting. References. 7 Using
Intramolecular Fluorescence Resonance Energy Transfer to Study Receptor
Conformation (Cornelius Krasel and Carsten Hoffmann). 7.1 Introduction. 7.2
Methods and Approaches. 7.3 Troubleshooting. References. 8 A Disulfide
Cross-linking Strategy Useful for Studying Ligand-induced Structural
Changes in GPCRs (Jian Hua Li, Stuart D.C. Ward, Sung-Jun Han, Fadi F.
Hamdan and Jürgen Wess). 8.1 Introduction. 8.2 Methods and Approaches. 8.3
General Considerations, Caveats and Troubleshooting. Acknowledgements.
References. 9 Use of Fluorescence Correlation Spectroscopy to Study the
Diffusion of G-protein-coupled Receptors (Stephen J. Briddon, Jonathan A.
Hern and Stephen J. Hill). 9.1 Introduction. 9.2 Methods and Approaches.
9.3 Troubleshooting. References. 10 Identification and Proteomic Analysis
of GPCR Phosphorylation (Kok Choi Kong, Sharad C. Mistry and Andrew B.
Tobin). 10.1 Introduction. 10.2 Methods. Acknowledgements. References. 11
Measurement and Visualization of G-Protein-coupled Receptor Trafficking by
Enzyme-linked Immunosorbent Assay and Immunofluorescence (Stuart J.
Mundell, Shaista P. Nisar and Eamonn Kelly). 11.1 Introduction. 11.2
Methods and Approaches. 11.3 Troubleshooting. References. 12 Substituted
Cysteine Accessibility Method (SCAM) (George Liapakis and Jonathan A.
Javitch). 12.1 Introduction. 12.2 Methods and Approaches. 12.3
Troubleshooting. References. 13 Homology Modelling of G-Protein-coupled
Receptors (John Simms). 13.1 Introduction. 13.2 Methods and Approaches.
13.3 Troubleshooting. 13.4 Automated Methods for Generating Models of
GPCRs. References. Appendix: Site-directed Mutagenesis and Chimeras (Alex
Conner, Mark Wheatley and David Poyner). A.1 Introduction. A.2 Why
Mutagenesis? A.3 Troubleshooting. A.4 Conclusion. References. Index.