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Multicolor Fiber FISH

Various genetic abnormalities can be observed in many human diseases. They vary from numerical chromosomal aberrations, deletions, and amplification of specific regions to chromosomal translocations and insertions. In the past decade, numerous fluorescence in situ hybridi ...

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Multi-Telomere FISH

The standard investigation for suspected chromosomal rearrangements in patients is cytogenetic analysis at a 400–550 band resolution, yet this cannot routinely detect rearrangements smaller than 5 Megabases (Mb), and much larger abnormalities escape notice if they occur in regi ...

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Fluorescence Genotyping for Screening Cryptic Telomeric Rearrangements

Mental retardation (MR), defined as an intelligence quotient (IQ) less than 70, represents the most frequent serious handicap in children and young adults. Moderate to severe MR (IQ

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Microarray CGH

Comparative genomic hybridization (CGH) to metaphase chromosome targets (1,2) has significantly contributed to our understanding of the cancer cytogenetics of more complex malignancies such as solid tumors (3,4). This molecular cytogenetics-based technique (hereafter r ...

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Chromosome Microdissection

It has been known for decades that chromosome rearrangements exist in most if not all human tumors (1) and certain human hereditary diseases (2). Distinct chromosomal abnormalities in tumors lead to the activation of proto oncogene products, creation of tumor specific fusion proteins, or i ...

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Primed In Situ Labeling

Prins (Primed in situ labeling) produces the same type of result as FISH; namely a hybridization dependant staining of specific DNA sequences in cell and tissue preparations. However, the staining is obtained differently, as the probe used in PRINS is unlabeled and staining results from the sy ...

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Spectral Karyotyping

Historically in clinical cytogenetics, G-banding has been the gold standard for detecting gross chromosomal abnormalities, ranging from simple numerical changes to the identification of complex structural rearrangements in clinical samples. The designation “marker chr ...

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Microarray Analysis of Gene Expression in Murine Cardiac Graft Infiltrating Cells

Microarray technology can rapidly generate large databases of gene expression profiles. Our laboratory has applied these techniques to analyze differential gene expression in cardiac tissue and cells based on mouse heart transplantation. We have analyzed the different gene exp ...

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Expression Profiling Using Affymetrix GeneChip Probe Arrays

Large-scale microarray expression profiling studies have helped us to understand basic biological processes and to classify and predict the prognosis of cancers; they have also accelerated the identification of new drug targets. Affymetrix GeneChip� probe arrays are high-dens ...

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Serial Analysis of Gene Expression (SAGE): A Useful Tool to Analyze the Cardiac Transcriptome

Serial analysis of gene expression (SAGE), a functional genomics technique, can be used for global profiling of gene transcripts. It relies on the preparation and sequencing of cDNA concatemers, but it does not require prior knowledge of the genes to be assayed (as with microarrays). Once analy ...

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Functional Genomics by cDNA Subtractive Hybridization

The regulation of myocardial gene expression is highly sensitive to any extracellular or intracellular stimulus that affects contractile function. Subtractive suppression hybridization represents a large-scale, unbiased method for detecting transcriptionally and ...

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Statistical Methods in Cardiac Gene Expression Profiling: From Image to Function

By providing genome-scale information on gene expression, microarray technology has gained popularity in diverse areas including clinical medicine. However, the analysis and interpretation of microarray data are often complicated. This chapter describes various strate ...

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Measurement of Cardiac Gene Expression by Reverse Transcription Polymerase Chain Reaction (RT-PCR)

Two methods applicable to the measurement of cardiac gene expression by reverse transcription polymerase chain reaction (RT-PCR) are presented. Each method describes a specific technique and includes information on how to optimize the experiments and how to trouble-shoot any prob ...

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Quantitative (Real-Time) RT-PCR in Cardiovascular Research

Quantitative (real-time) PCR (qPCR) represents a highly sensitive, sequence-specific, and reproducible technique for the gel-free detection and quantitation of nucleic acids. Owing to its large dynamic range and throughput, this approach has become the chosen method for rapid qua ...

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RNase Protection Assay for Quantifying Gene Expression Levels

Quantifying the level of mRNA is central to the study of mammalian gene expression. Conventional approaches such as Northern blotting are often prone to low sensitivity and reproducibility. The RNase protection assay (RPA) provides a sensitive alternative for the detection and quant ...

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Characterization of cis-Regulatory Elements and Transcription Factor Binding: Gel Mobility Shift Assay

To understand how cardiac gene expression is regulated, the identification and characterization of cis-regulatory elements and their trans-acting factors by gel mobility shift assay (GMSA) or gel retardation assay are essential and common steps. In addition to providing a general p ...

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In Situ Hybridization: A Technique to Study Localization of Cardiac Gene Expression

In situ hybridization allows the detection of specific gene transcripts in tissues, cells or, chromosomes. In the cardiovascular field, this powerful and rapid methodology provides precious insights into the complex gene organization and expression within an heterogeneous cell ...

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Promoters to Express Cloned Genes Uniformly in Drosophila

Forcing ubiquitous expression of a given gene in transgenic Drosophila has become a powerful tool of molecular genetics. Gene misexpression usually causes a dominant phenotype and one may deduce potential functions for the gene from an analysis of this phenotype. Evidently, when a domin ...

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Strategies in Generating Transgenic Mammals

The ability to manipulate genes in mammals is providing insights into most aspects of modern biology, including the regulation and function of genes, the mechanisms of developmental and pathological processes, and the generation of animal models for human disorders. Furthermore, the ...

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Application of Micromechanical Piercing Structures for Genetic Transformation of Nematodes

Several methods are available to accomplish gene transfer into mammalian cells, plant cells, yeast, and other organisms. Three of the most promising DNA delivery systems are electroporation (1,2), DNA-coated microprojectiles (3), and microinjection (4,5). Electroporation invo ...

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