This book describes how to perform and optimize the various types of Polymerase Chain Reactions (PCR) for postgraduate students, scholars and researchers in all branches of life science. PCR is a method widely used to rapidly make millions to billions of copies of specific DNA samples, allowing scientists to take a very small sample of DNA and amplify it (or a part of it) to a large enough amount to study in detail. This book also deals with molecular biology reagents preparation and general laboratory procedures, equipment use and safety precautions. The various forms of pathogenic agents drastically affect human society and bring human life notoriously. The correct and exact details of these creatures can be derived through the prompt diagnosis of pathogens as early as possible. The current form of diagnosis is molecular diagnostics, but optimization and standardization are most important for the exact quality of results. This book is written with the need to address the technical problems while optimizing the PCR reactions in mind. The same procedure is fully applicable whenever techniques are being handled in life science laboratories. The textbook encourages the persons who engage in microbiology, molecular biology and life science laboratory to accept and implement basic concepts in various types of PCRs and develop in-house techniques for day-to-day routine activities. This book also deals with the major junk areas while designing primer for various types of PCRs and deals with how to address and troubleshoot the issues that arise while doing various forms of PCRs. This book also deals with post-PCR activities and troubleshooting of gel electrophoresis

James D. Watson When, in late March of 1953, Francis Crick and I came to write the first Nature paper describing the double helical structure of the DNA molecule, Francis had wanted to include a lengthy discussion of the genetic implications of a molecule whose struc ture we had divined from a minimum of experimental data and on theoretical argu ments based on physical principles. But I felt that this might be tempting fate, given that we had not yet seen the detailed evidence from King's College. Nevertheless, we reached a compromise and decided to include a sentence that pointed to the biological significance of the molecule's key feature-the complementary pairing of the bases. "It has not escaped our notice," Francis wrote, "that the specific pairing that we have postulated immediately suggests a possible copying mechanism for the genetic material." By May, when we were writing the second Nature paper, I was more confident that the proposed structure was at the very least substantially correct, so that this second paper contains a discussion of molecular self-duplication using templates or molds. We pointed out that, as a consequence of base pairing, a DNA molecule has two chains that are complementary to each other. Each chain could then act ". . . as a template for the formation on itself of a new companion chain, so that eventually we shall have two pairs of chains, where we only had one before" and, moreover, " ...

Do you want to know the details that should be taken into consideration in order to have accurate conventional and real-time PCR results? If so, this book is for you. Polymerase Chain Reaction for Biomedical Applications is a collection of chapters for both novice and experienced scientists and technologists aiming to address obtaining an optimized real-time PCR result, simultaneous processing of a large number of samples and assays, performing PCR and RT-PCR on cell lysate without extraction of DNA or RNA, detecting false-positive PCR results, detecting organisms in viral and microbial diseases and hospital environment, following safety assessments of food products, and using PCR for introduction of mutations. This is a must-have book for any PCR laboratory.

The polymerase chain reaction (PCR) is the most sensitive method for revealing the presence of otherwise undetectable quantities of the genome of RNA or DNA of human viruses. The Polymerase Chain Reaction (PCR) for Human Viral Diagnosis addresses the urgent need to use this revolutionary technology in reference and routine diagnostic laboratories. It informs the molecular biologist of the most appropriate clinical uses for PCR and educates the clinician and medical virologist about the subtleties and benefits of gene amplification. The reader is given an understanding and appreciation of the principles of PCR and how, why, and where it should be applied. The book explains the principles behind PCR and its role in the diagnostic and public health laboratory. The application of PCR to the detection and investigation of viral latency and persistence is presented by the originators of in situ amplification. There are individual contributions from experts in their respective fields on the detection, characterization, and analysis by PCR of gastroenteritis viruses, hepatitis viruses, herpesviruses, rhinoviruses, enteroviruses, flaviviruses, polyomaviruses, human immunodeficiency virus (HIV), human T-lymphotropic virus types I and II (HTLV-I and II); and measles, mumps, rubella, influenza, rabies, and B19 viruses.

This book is intended to present current concepts in molecular biology with the emphasis on the application to animal, plant and human pathology, in various aspects such as etiology, diagnosis, prognosis, treatment and prevention of diseases as well as the use of these methodologies in understanding the pathophysiology of various diseases that affect living beings.

This PCR handbook establishes an easy approach for the students going through their studies in Molecular Diagnostics. It introduces timely topics such as Clinical Applications of PCR, Real time PCR, PCR Optimization, Application of PCR, PCR Array System Performance, Designing the primers, PCR in diagnosis of diseases and different PCR Techniques and their procedures.

In this laboratory "cook-book", the authors provide a concise guide to PCR-based techniques to quantify nucleic acids in biological and clinical samples using exclusively nonradioactive detection methods, e.g. HPLC, biotin and digoxigenin based protocols. Each method presentation also includes sections on theory, reagents, standards, applicability, limitations, and trouble shooting. In addition to the protocols, the authors also provide the necessary information on: general aspects of nucleic acid quantitation; design of PCR standards; mRNA purification; cDNA synthesis; solution hybridization; DNA sequencing. This laboratory guide enables professionals as well as beginners to adopt easily quantitative PCR protocols into their own clinical or biomedical research.

The basis for the effective treatment and cure of a patient is the rapid diagnosis of the disease and its causative agent, which is based on the analysis of the clinical symptoms coupled with laboratory tests. Although rapid advance ments have been made in the laboratory diagnosis of virus diseases, the neces sary isolation of the causative virus from the clinical specimens is a relatively long procedure. Viruses which integrate into the cellular DNA (such as human immunodeficiency virus, HIV -1, or hepatitis B virus) are difficult to identify by molecular techniques, while viruses which exist in the clinical material in low concentrations are even more formidable to identify. Recently, the application of the polymerase chain reaction (peR) technique developed by K. D. Mullis and detailed in the study by Saiki et al. (1985) led to a revolution in virus diagnosis. The peR technique was rapidly applied to the diagnosis of viruses in clinical material. Volume 1 of Frontiers of Virology provides new information on the advan tages of the use of the peR for the diagnosis of many human disease-causing viruses, as well as on some problems with its use.