As our understanding of molecular biology continues to develop, both general improvements and specialized tools are needed for the progression of RNA detection and analysis. This thesis describes three distinct projects within the broad goal of expanding the techniques available for better interrogation of RNA. Chapter II illustrates the development of a new technique for microRNA detection. MicroRNAs are a class of small regulatory RNAs present in eukaryotes. MicroRNAs pose a challenge for detection because of their small size and sequence homology, but have drawn increasing interest due the predicted prevalence of their gene targets and links between their expression levels and cancer. The development of an isothermal detection strategy is described which incorporates two nucleic acid templated steps, two amplification steps, and fluorescent detection. Up to 3000-fold amplification can be obtained, and 200 pM quantities of let-7a microRNA and above can be detected reliably. In Chapter III, two distinct approaches are taken for the detection of N(6)-methyladenosine, the most common modification in RNA: harnessing base-pairing selectivity, and developing selective reactions. A successful method was developed that harnessed the inherently higher efficiency of a commercially-available polymerase, Tth DNA polymerase, when pairing T with adenosine as opposed to N(6)-methyladenosine. Kinetic analysis showed that the selectivity of the enzyme ranged from 4-18-fold, depending on sequence context. Detection of m6A in ribosomal RNA and in a highly-expressed messenger RNA are demonstrated. The final chapter focuses on a mechanistic approach for enhancing RNA recovery after formaldehyde treatment using an organocatalyst. Mass spectrometry and gel electrophoresis showed an enhanced rate of adduct reversal in the presence of the catalyst, and effects of temperature, buffer, and pH were also analyzed. The results from monomeric and oligomeric test systems have subsequently been applied to recovery of RNA from fixed samples, showing ~2-fold enhancement in recovered RNA due to the catalyst and up to 25-fold enhancement when the catalyst was used in conjunction with optimized time and temperature.

This laboratory guide represents a growing collection of tried, tested and optimized laboratory protocols for the isolation and characterization of eukaryotic RNA, with lesser emphasis on the characterization of prokaryotic transcripts. Collectively the chapters work together to embellish the RNA story, each presenting clear take-home lessons, liberally incorporating flow charts, tables and graphs to facilitate learning and assist in the planning and implementation phases of a project.RNA Methodologies, 3rd edition includes approximately 30% new material, including chapters on the more recent technologies of RNA interference including: RNAi; Microarrays; Bioinformatics. It also includes new sections on: new and improved RT-PCR techniques; innovative 5' and 3' RACE techniques; subtractive PCR methods; methods for improving cDNA synthesis.* Author is a well-recognized expert in the field of RNA experimentation and founded Exon-Intron, a well-known biotechnology educational workshop center * Includes classic and contemporary techniques * Incorporates flow charts, tables, and graphs to facilitate learning and assist in the planning phases of projects

Viral respiratory tract infections are important and common causes of morbidity and mortality worldwide. In the past two decades, several novel viral respiratory infections have emerged with epidemic potential that threaten global health security. This Monograph aims to provide an up-to-date and comprehensive overview of severe acute respiratory syndrome, Middle East respiratory syndrome and other viral respiratory infections, including seasonal influenza, avian influenza, respiratory syncytial virus and human rhinovirus, through six chapters written by authoritative experts from around the globe.

Scores of talented and dedicated people serve the forensic science community, performing vitally important work. However, they are often constrained by lack of adequate resources, sound policies, and national support. It is clear that change and advancements, both systematic and scientific, are needed in a number of forensic science disciplines to ensure the reliability of work, establish enforceable standards, and promote best practices with consistent application. Strengthening Forensic Science in the United States: A Path Forward provides a detailed plan for addressing these needs and suggests the creation of a new government entity, the National Institute of Forensic Science, to establish and enforce standards within the forensic science community. The benefits of improving and regulating the forensic science disciplines are clear: assisting law enforcement officials, enhancing homeland security, and reducing the risk of wrongful conviction and exoneration. Strengthening Forensic Science in the United States gives a full account of what is needed to advance the forensic science disciplines, including upgrading of systems and organizational structures, better training, widespread adoption of uniform and enforceable best practices, and mandatory certification and accreditation programs. While this book provides an essential call-to-action for congress and policy makers, it also serves as a vital tool for law enforcement agencies, criminal prosecutors and attorneys, and forensic science educators.

This edited book, "Nucleic Acids - From Basic Aspects to Laboratory Tools", contains a series of chapters that highlight the development and status of the various aspects of the nucleic acids related to DNA chemistry and biology and the molecular application of these small DNA molecules and related synthetic analogues within biological systems. Furthermore, it is hoped that the information in the present book will be of value to those directly engaged in the handling and use of nucleic acids, and that this book will continue to meet the expectations and needs of all who are interested in the different fascinating aspects of molecular biology.

2775 references to research projects being conducted in the United States and elsewhere. Entries arranged under 11 topics, e.g., Cancer therapy, Supportive care of cancer patients, and Rehabilitation. Entries include title, researcher, address, contract number, summary, and supporting agency. Indexes by subjects, investigators, contractors, supporting agencies, and contractor numbers.