遗传学实验技术

Immunocytochemistry and in situ hybridization—the in situ staining approaches for detection of antigens, DNA, or RNA in intact cells and tissue sections— are common and powerful tools for biological and biomedical research and clinical diagnostics. By combining the exponential am ...

Since the discovery of the Hepatitis C Virus (HCV) in 1989 (1), known to that point as the infectious agent of Non-A Non-B hepatitis (2), there has been an intensive focus on understanding the underlying biology of its disease process. Of significant interest has been the study of heterogenous populat ...

The hepatitis C virus (HCV) is a major cause for liver-related morbidity and mortality worldwide. Quantification of HCV-RNA has become an important tool to tailor and monitor antiviral therapies for this disease (1). Although a low viral load is associated with a higher efficacy of Interferonm ...

The use of molecular-based methodologies for the identification of viral nucleic acid has evolved into a clinically useful adjunct to the biochemical, serological, and histological assays used to evaluate patient health. The clinical application of viral-load assays for monitor ...

PCR (polymerase chain reaction)products are conventionally detected by agarose gel electrophoresis and ethidium bromide staining.However, the specificity and sensitivity of detection can be increased by hybridization with a labeled probe that is complementary to an inter ...

The quantitative measurement of specific mRNA species is of major importance for studies on gene expression. Northern blotting is a relatively insensitive method requiring microgram amounts of RNA. It is time consuming and semi-quantitative at best. Because of the limitations of Nort ...

The ability to detect mRNA in tissue sections provides a powerful tool for localizing gene expression in complex tissues comprising mixed cell populations. Detection of gene expression at the mRNA level is particularly important to confirm the identity of cells expressing soluble or se ...

It has long been questioned whether Mycobacterium paratuberculosis plays a role in the aetiology of Crohn’s disease (1,2). Several methods of detection (serology, culture, and PCR)have been employed to prove or disprove a link, but without broad consensus to date (3, 4). With granulomatous di ...

Laser-capture microdissection (LCM) is a powerful method for efficient procurement of defined cell populations from whole tissue sections. It enables the study of cell biology and molecular pathology at the level of DNA, RNA, and protein (1,2,3). The principle of LCM, as applied by the Pixcell sy ...

Sequential chromatin immunoprecipitation (SeqChIP) assays have been developed for the study of interactions of two or more proteins (or simultaneous histone modifications) at genomic sites. It is based on the principle that chromatin and associated proteins can be first immunopr ...

Quantitative polymerase chain reaction (Q-PCR) allows for the accurate and reproducible determination of the amount of target DNA in a sample through the measurement of PCR product accumulation in “real time.” This method determines starting target DNA quantity over a large assay dynam ...

Native chromatin immunoprecipitation refers to a method that allows for identification and quantification of DNA that is associated with specific chromatin proteins without altering the structure of these proteins. The method has been used with great success in the past and has some ad ...

Chromatin immunoprecipitation (ChIP) is a widely used technique to get a snap-shot of protein–DNA interactions in cells. ChIP has notably been used for mapping the location of modified histones, transcription factors, or chromatin remodeling enzymes in the genome, often in relation to t ...

DNA methyltransferases are important enzymes and their inhibition has many potential applications. The investigation of DNA methyltransferases as well as screening for potential inhibitors requires specialized enzyme assays. In this chapter, we describe three DNA methylt ...

DNA methylation plays a significant role in the expression of the genetic code and affects early growth and development through its influence on gene expression. DNA methyltransferase 1 (Dnmt1) is the enzyme responsible for maintaining the methylation marks through cell division. Ho ...

Epigenetic alterations regulate the utilization of the genome by permitting or inhibiting access of transcription factors and associated complexes. Although there are several different types of epigenetic alterations, such as acetylation and methylation of histone tails, t ...

Alterations in DNA methylation patterns are implicated in playing a major role in the development of cancer, thus highlighting the need to continually develop new technologies to analyze epigenetic marks. Methylated-CpG Island Recovery Assay (MIRA), based on the high affinity of the MB ...

Epigenetic mechanisms control gene transcription primarily through regulating chromatin structures and DNA methylation. Transcription factors can also affect gene transcription through binding of the key transcriptional machinery to the gene promoter. These factors n ...

Epigenetics refers to the collective heritable changes in phenotype that arise independent of genotype. Two broad areas of epigenetics are DNA methylation and histone modifications and numerous techniques have been invented to analyze epigenetic processes not only at the level of s ...

The crucial roles played by RNA-binding proteins in all aspects of RNA metabolism, particularly in the regulation of transcription, have become increasingly evident. Moreover, other factors that do not directly interact with RNA molecules can nevertheless function proximally to R ...