Preface

In this post-genomic era of modern biology, a large number of molecular biology manuals are available on the market. As the authors of this manual, we feel committed to addressing this logical question: “What is the need to write another molecular biology manual?” Although several excellent molecular biology manuals are available nowadays, most of these manuals are aimed at catering to the relatively trained and experienced workers/experimenters in the field. Notably, all of them extensively cover almost all the methodologies that are widely used not only in molecular cloning but also in wider areas of molecular biology, including genomic libraries, RNA and protein methodologies, molecular genetics, genomics, and proteomics. An apprentice in the field often gets lost when s/he is offered one such manual to consult for carrying out elementary experiments involving the isolation of genomic DNA from a common source organism or even carrying out a simple cloning experiment. Thus, we feel that, indeed, there is a genuine need for a manual that will cater to the essence of the key methodology involved in molecular cloning and help the relative newcomers in the field.

In composing the chapters in this book, we deliberately remain “cloning-centric” and focus on appending only those methods that are directly associated with molecular cloning processes and stay away from methods covering diverse areas of modern biology. We feel that the selection of the methods will make the manual simple and succinct, which will help the new students learn about a specific/selected method more easily. Given the risk of handling various laboratory elements, including chemical, biological, and radiological reagents, as well as safety precautions to deal with them, an introductory chapter addressing these issues is included. This chapter also provides various types of fundamental skills needed to train an individual adequately into an experienced experimenter. Considering the fact that during this training, a beginner must learn sufficient background of a given method, we provide sound explanations of rationale and modes of action for all individual steps. Moreover, after specific individual steps in all the protocols, we provide notes (presented in italics) that cover many critical parameters, modes of action, and do’s and don’ts involved in that step, which we think will be very helpful for the new students to master the entire method. Furthermore, to provide sufficient background information, we present special information panels on selected topics, reagents, methods, etc., in every chapter that would equip an apprentice with profound knowledge and information on various topics associated with molecular cloning. Finally, we included an appendix at the end that lists the composition and methods of preparation of all the reagents/buffers/solutions used in diverse protocols and the most commonly used methods that are involved in various techniques.

Finally, we would like to thank our students who provided various feedback and several sample photographs from their experiments that were used in different chapters in this manual. Last but not least, we would like to apologize for any undetected and inadvertent mistakes/errors that might still be present despite a rigorous attempt to eliminate them.