This chapter describes a method for the multiplex analysis of six biallelic single nucleotide polymorphisms (SNPs) associated with thrombophilia. The method may, however, be adapted for the simultaneous analysis of up to 100 markers (50 biallelic SNPs) in a single reaction. In the method de ...
Class I and II loci of the human leukocyte antigens (HLA) represent the most polymorphic region of the genome. Evolutionary pressure has resulted in a large number of allelic variants of these loci ensuring the high frequency of heterozygous genotypes observed in human populations. Molecu ...
Inflammation is becoming increasingly recognized and discussed as an important pathobiochemical origin in many disease entities such as atherosclerosis, cancer, or infections and genetically determined susceptibility to danger signals may influence the development of i ...
One major goal of genetic research is to understand the role of genetic variation in living systems. In humans, by far the most common type of such variation involves differences in single DNA nucleotides, and is thus termed single nucleotide polymorphism (SNP). The need for improvement in throu ...
The increasing emphasis in life science research on utilization of genetic and genomic information underlies the need for high-throughput technologies capable of analyzing the expression of multiple genes or the presence of informative single nucleotide polymorphisms (SNPs) ...
Gene expression profiling of peripheral blood is a very attractive approach for the development of new diagnostic markers of blood-borne but also systemic diseases as well as the development of biomarkers for drug development. Since most cellular components of peripheral blood are spe ...
The differential display-polymerase chain reaction (DD-PCR) technique is a unique, sequence independent tool for mRNA profiling and relative quantification. It is particularly suited for clinical samples yielding limited amounts of RNA. Unlike closed systems like microarra ...
The comparison of two RNA populations that differ from the effects of a single-independent variable, such as a drug treatment or a specific genetic defect, can identify differences in the abundance of specific transcripts that vary in a population-dependent manner. There are a variety of met ...
Platelets are anucleated cells that are generated from megakaryocytes via thrombopoiesis. They lack genomic DNA but have a pool of individual mRNA transcripts. Taken together, these mRNAs constitute a platelet transcriptome. Platelets have a unique and reproducible transcript p ...
Human blood platelets are anucleate cells that contain minute amounts of translational active mRNA. Investigation of the gene expression profile by microarray analysis has become an excellent tool for better understanding of normal and pathological platelet function. Its use, ho ...
Platelet transcriptome studies provide a powerful tool in the analysis of disorders affecting the megakaryocytic-platelet lineage. However, individualised platelet gene expression profiling is hampered by the exceptionally low yield of platelet mRNA. The yield of mRNA trans ...
MiRNAs are non-coding RNAs of 19–25 nucleotides in length that regulate gene expression. Recent work suggests a role for miRNAs during hematopoiesis. Here, we described in detail how to successfully obtain highly pure megakaryocytes precursors from human CD34+ stem cells in culture to per ...
Reticulocytes are the last stage of erythropoiesis before red blood cells (RBC). Although most of the RBC proteins have been characterized, little is known about expression profile of their mRNA during differentiation. Our goal was to initiate a genomic global solution to provide a transcr ...
Induction of an immune response to a particular antigen is the basis of vaccination. This has been done for years to prevent infectious diseases, and has the potential for the treatment of cancer. The immune response is nowadays more precisely modulated rather than simply induced, like in case of im ...
We here describe the methods for the isolation of distinct hematopoietic subpopulations, as defined by their immune phenotype by fluorescence-activated cell sorting, and how these cells can be analyzed even at a single-cell level for the gene expression of a number of transcription facto ...
All cellular components of the blood originate from hematopoietic stem cells in the bone marrow. These pluripotent stem cells give rise to two lineages of specialized cells (Fig. 1). The lymphoid lineage produces B- and T-cells; the myeloid lineage gives rise to leukocytes, erythroblasts (the ...
Traditionally, studies on the development of atherosclerosis have been performed in animals larger than mice, with a particular preponderance of studies in rabbits. Studies in the mid-1980s began to introduce the mouse as a model for the development of atherosclerosis. The extensive ge ...
The laboratory mouse offers tremendous opportunities for studies on the genetic basis of cancer. No other model system is so easily and widely used to test the effect of potentially oncogenic mutations in a living animal. Mouse strains differ extraordinarily in their innate susceptibilty ...
Recently, transgenic technologies to express cloned genes in the mouse have become fully mature. These advances have made the mouse one of the most useful animal models for biomedical research. The main strength of the mouse, compared with other model organisms, such as worms (Caenorhabdit ...
The Fragment Assembly System is a series of related programs that help you assemble overlapping fragment sequences as obtained at the lab bench. Specifically, the programs resemble a small databas system that helps you to achieve six major tasks:
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