Gene Quantification

Geneticists and molecular biologists have been interested in quantifying genes and their products for many years and for various reasons (Bishop, 1974). Early molecular methods were based on molecular hybridization, and were devised shortly after Marmur and Doty (1961) first showed that denaturation of the double helix could be reversed - that the process of molecular reassociation was exquisitely sequence dependent. Gillespie and Spiegelman (1965) developed a way of using the method to titrate the number of copies of a probe within a target sequence in which the target sequence was fixed to a membrane support prior to hybridization with the probe - typically a RNA. Thus, this was a precursor to many of the methods still in use, and indeed under development, today. Early examples of the application of these methods included the measurement of the copy numbers in gene families such as the ribosomal genes and the immunoglo bulin family. Amplification of genes in tumors and in response to drug treatment was discovered by this method. In the same period, methods were invented for estimating gene num bers based on the kinetics of the reassociation process - the so-called Cot analysis. This method, which exploits the dependence of the rate of reassociation on the concentration of the two strands, revealed the presence of repeated sequences in the DNA of higher eukaryotes (Britten and Kohne, 1968). An adaptation to RNA, Rot analysis (Melli and Bishop, 1969), was used to measure the abundance of RNAs in a mixed population.

This volume provides experimental and bioinformatics approaches related to different aspects of gene expression analysis. Divided in three sections chapters detail wet-lab protocols, bioinformatics approaches, single-cell gene expression, highly multiplexed amplicon sequencing, multi-omics techniques, and targeted sequencing. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Gene Expression Analysis: Methods and Protocols aims provide useful information to researchers worldwide.

Can the son or daughter of a baseball pitcher or cricket bowler throw a ball 100 miles an hour? Is the son or daughter of an opera singer also an opera singer? Is a house with functional light switches lit? The line of thinking in these rhetorical questions also applies to human genetics. What do baseball pitchers, opera sing ers, light switches, and the Human Genome Project have in common? These questions address the issue of potential versus realization of function. Although sons and daughters of baseball pitchers and opera singers may have inherited the mechanical attributes to be baseball pitchers and opera singers, they may not, at any point in time, be baseball pitchers or opera singers. A house with functional light switches is not lit unless the light switches are on. Similarly, all of the genes discovered and sequenced as a result of the Human Genome Project are not expressed at the same time. Genome project information will allow us to deter mine the repertoire of genes in an individual, which is analogous to determining where the light switches in a house are located and whether they are functional (a mutation or deletion in the Genome Project Model). The pattern of "on" light switches in a house gives us functional information as to what the family inside is doing (e. g. , eating, reading, sleeping). Similarly, the pattern of gene expression (RNA) gives us information on what our bodies are doing (e. g.

With the advent of high-throughput technologies following completion of the human genome project and similar projects, the number of genes of interest has expanded and the traditional methods for gene function analysis cannot achieve the throughput necessary for large-scale exploration. This book brings together a number of recently developed techniques for looking at gene function, including computational, biochemical and biological methods and protocols.

Edited by two leaders, this book has drawn together expertise from around the globe to form a unified, cohesive resource for the droplet microfluidics community. Starting with the basic theory of droplet microfluidics before introducing its use as a tool, the reader is treated to chapters on important techniques, including robust passive and active droplet manipulations and applications such as single cell analysis, which is key for drug discovery. This book is a go-to resource for the community yearning to adopt and promote droplet microfluidics into different applications.

This book presents practical approaches for the analysis of data from gene expression micro-arrays. It describes the conceptual and methodological underpinning for a statistical tool and its implementation in software. The book includes coverage of various packages that are part of the Bioconductor project and several related R tools. The materials presented cover a range of software tools designed for varied audiences.

Focusing on Saccharomyces cerevisiae, the second edition of Yeast Gene Analysis represents a major reworking of the original edition, with many completely new chapters and major revisions to all previous chapters. Originally published shortly after completion of the yeast genome sequence, the new edition covers many of the major genome-wide strategies that have been developed since then such as microarray analysis of transcription, synthetic gene array studies, protein microarrays and chemical genetic approaches. It represents a valuable resource for any research laboratory using budding yeast as their experimental system in which to identify new yeast gene functions. The chapters are written in a readable style with useful background information, technical tips and specific experimental protocols included as appropriate, enabling both the novice and the experienced yeast researcher to adopt new procedures with confidence. New chapters on: Strain construction; genome-wide two-hybrid approaches; use of microarrays for transcript analysis; real-time analysis of chromosome behaviour and FRET; synthetic gene array technology and protein arrays; chemical genomics and yeast prions; RNA gene analysis and mitochondrial gene function analysis; phylogenetic footprinting; discovering human gene function and predicting yeast gene function