基因表达差异显示实验

The introduction of primed in situ labeling (PRINS) into plant cytogenetics provided a novel means for fast and highly specific visualization of DNA sequences on chromosomes and in interphase nuclei. Although the technique does not reach the sensitivity of fluorescence in situ hybridi ...

The preparation of a somatic cell tissue is a key point of any successful molecular biology and genetic analysis of chromosome complement and genome variations. Current molecular biology investigations require an increased amount of different tissues to study. The genetic studies of t ...

Fetal nucleated cells circulating in the peripheral blood during pregnancy are potential targets for noninvasive genetic testing. Fluorescence in situ hybridization (FISH) frequently is used to quantify the total number of fetal cells in peripheral blood of pregnant women. We desc ...

In situ amplification techniques are designed to increase the mass of DNA in a fixed target, either whole cells or tissue sections. When combined with fluorescently labeled nucleotides, they can be used for locus detection. They also can be used to increase target mass for subsequent operation ...

An ultrarapid three- and four-color primed in situ (PRINS) procedure has been developed for rapid chromosome identification and aneuploidy assessment on isolated cells. Based on the direct in situ mixing of fluorochromes (fluorescein isothiocyanate, tetramethylrhodamine i ...

It is now well recognized that chromosomal translocation followed by overexpression of a chimeric gene product plays a critical role in tumorigenicity in various malignant tumors, especially those of leukemia, malignant lymphoma, and soft-tissue tumors. In these malignant tumors, ...

In situ detection techniques allow specific nucleic acid sequences to be exposed in morphologically preserved tissue sections. In combination with immunocytochemistry, in situ detection can relate microscopic topological information to gene activity at the transcript or pr ...

Cytomegalovirus latency depends on an interaction with hematopoietic cells in bone marrow and peripheral blood. The distribution of latent viral DNA and transcripts in these cells was investigated using methods based on polymerase chain reaction (PCR)-driven in situ hybridizat ...

Since the first publication on the method of in situ polymerase chain reaction (PCR), several thousand research papers have appeared in peer-reviewed journals describing various findings based solely on the application of this method or combined with other more robust methods, includ ...

The advent of molecular genetic techniques has brought forth new procedures for in situ chromosomal analysis. One of these techniques is the primed in situ labeling (PRINS) procedure, which constitutes a fast and efficient alternative to conventional fluorescence in situ hybridiza ...

Micronuclei are indirect but reliable indicators of chromosome damage, formed at the mitotic division after either chromosome loss or chromosome breakage events, and identified at interphase as small bodies of extra-chromatin in the cytoplasm of mammalian cells. In this chapter, the ...

The protocol is suited for the quantitative and qualitative detection of simple repeat target DNA composed of three or fewer of the four bases A, C, G, and T. A consequence of the lacking base(s) is that such DNA can be synthesized from nucleotide mixtures containing the particular bases as dideoxynucl ...

An efficient RNAi largely depends on optimal design of the siRNA. In recent studies, Dicer substrates were found to be more potent than classical synthetic 21-mer siRNAs, suggesting a coupling of the Dicer-mediated processing step to the efficient assembly of the silencing complex, RISC. We d ...

RNA interference (RNAi) is a form of posttranscriptional gene silencing mediated by microRNA (miRNA) and small interfering RNA (siRNA). In Drosophila melanogaster, the RNase III enzymes Dicer-1 and Dicer-2 generate miRNA and siRNA, respectively. We describe the methods for the expres ...

Recent studies have revealed that Argonaute proteins are crucial components of the RNA-induced silencing complexes (RISCs) that direct both small interfering RNA (siRNA)- and microRNA (miRNA)-mediated gene silencing. Full complementarity between the small RNA and its target me ...

RNA interference (RNAi) is an endogenous gene regulatory pathway that the research community has adopted to facilitate the creation of a functional map of the human genome. To achieve this, small interfering RNAs (siRNAs), short synthetic duplexes having complete homology to the intend ...

RNA-mediated interference (RNAi) has been a valuable tool for the analysis of gene function in Caenorhabditis elegans (C. elegans). In C. elegans, the injection of double-stranded RNA (dsRNA) or plasmid DNA expressing dsRNA under the control of a C. elegans promoter results in gene inactivati ...

Containment of the SARS coronavirus (SCV) outbreak was accompanied by the rapid characterization of this new pathogen's genome sequence in 2003, encouraging the development of anti-SCV therapeutics using short interfering RNA (siRNA) inhibitors. A pair of siRNA duplexes identifi ...

We describe the protocols of using siRNAs, or shRNAs delivered by a lentiviral vector, as a means to silence cancer-causing genes. We use cervical cancer as a model to demonstrate the inhibition of the human papillomavirus (HPV) oncogenes E6 and E7 in cervical cancer cells by RNAi and inhibition of the ...

RNA interference (RNAi) is a sequence-specific gene regulatory mechanism in which the specificity is determined by small RNAs. Three major classes of endogenous small RNAs, namely microRNAs (miRNAs), small interfering RNAs (siRNAs), and piwi-interacting RNAs (piRNAs/gsRNAs), ha ...