PCR GURU - An Ultimate Benchtop Reference for Molecular Biologists

This 62-page eBook is an inspiring reference for PCR setup, optimization and troubleshooting. In the Background chapter, a brief history and the basic science underlying PCR is outlined. Procedure chapter starts with teaching how to accurately and successfully design primers and continues with detailed descriptions of standard PCR protocols. Various modifications of the standard protocols and steps for PCR optimization are described in the Troubleshooting chapter. Author's experience with PCR of about 8 years is distilled into Good Practices and Practical Tips chapters. The Special Cases chapter is describing several special applications of PCR and their optimization and troubleshooting. One of your favourite parts of this eBook will be the Appencices, which are listing many useful tables, figures and key protocols.

 




PCR Guru Cover      

 

Table of Contents

Preface
Chapter 1: Introduction
      1.1. A Bit of History
      1.2. What is PCR?
      1.3. PCR Components
      1.4. Exponential nature of PCR
      1.5. Popularity of PCR
Chapter 2: Procedure
      2.1. Primer Design
      2.2. Standard PCR setup
      2.3. Controls
      2.4. Cycling Parameters
Chapter 3: Good PCR Practices
Chapter 4: Troubleshooting PCR Problems

      4.1 Optimization rules
      4.2 General Optimization Strategies
          4.2.1. Annealing temperature gradient
          4.2.2. Mg++ Titration
          4.2.3. Hot-start
          4.2.4. Touchdown
          4.2.5. Additives
      4.3 Troubleshooting Specific Problems
          4.3.1. Problem type 1: No PCR products were observed.
          4.3.2. Problem type 2: Multiple PCR products were observed.
          4.3.3. Problem type 3: Smears were observed.
          4.3.4. Problem type 4: Observed bands were faint (low yield).
          4.3.5. Problem type 5: Bands observed in controls.
          4.3.6. Other Problem Types.
Chapter 5: Practical Tips
Chapter 6: Special Cases

      6.1. Long-PCR
      6.2. RAPD-PCR
      6.3. Multiplex PCR
Appendix A: Agarose Gel Electrophoresis Protocol
Appendix B: Silver Staining of DNA Gels
Appendix C: Plasmid Miniprep Protocol
Appendix D: Restriction Enzyme Digestion Protocol
Appendix E: Protocol for DNA Concentration Estimation by using Hoechst 33258
Appendix F: Formulas
Appendix G: Primers for Common Promoters, Housekeeping Genes, Reporters.
Appendix H: Tags
Appendix I: Thermostable DNA polymerases
Appendix J: Example of a Daily PCR Log
Appendix K: Common Buffers and Solutions
Appendix L: Useful numbers
Appendix M: Yeast colony PCR
Useful websites
Glossary and Abbreviations
Index

 


eBook Details
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Feedback from our customers

"Thank you very much for providing such an excellent introductory guide. Many successes with this first and subsequent editions. Kind regards, Frank Merante"

"I have purchased your book PCR-guru and it has been of immense help. Now people come to me with their doubts! :) Anyway, I was wondering if you have/are writing a book on cloning as well. When can we get more books like PCRguru?" - K.S.

"Your book is gorgeous!" - S.Z.



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About the author

PCR Guru was written by Ayaz Najafov, PhD. He is currently a postdoctoral researcher at Harvard Medical School, Cell Biology Department.

 

 


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