遗传学实验技术

The broad application of recombinant adenoviruses to the development of vaccines and gene therapy vectors has encouraged the development of molecular assays for the facile quantitation of adenoviral particles and the assignment of their infectious potency. The Genome Quantita ...

Virus-mediated gene transfer shows great potential as a therapeutic strategy for the management of various inherited and acquired human diseases. Among the current viral vectors, adeno-associated virus (AAV) has become the vector of choice for numerous gene therapy applications. As ...

Currently, with the exception of naked DNA formulations, most pharmaceutical preparations of plasmid DNA employ some type of polycationic delivery vector such as synthetic cationic polymers and lipids to enhance delivery. A number of biophysical techniques are readily available ...

The past 10 years have witnessed an increasing application of in situ hybridization techniques in biomedical research. This is mainly a result of the development of nonradioisotopic nucleic acid hybridization detection formats. Such methods allow much better exploitation of the sp ...

Different strategies have been introduced to facilitate characterization of complex genomes; the techniques include molecular cloning, agarose gel electrophoresis and blot-hybridization analysis, DNA sequencing, construction of somatic cell hybrid lines, in situ hybr ...

In preparations of highly condensed metaphase chromosomes, it is often difficult to resolve in situ hybridization signals of probes to nearby regions. This is in part, a result of traditional fixation procedures that use alcohol, acetic acid, and/or relatively strong ionic conditions, a ...

An innovative variation of fluorescence in situ hybridization (FISH) allows preparation of single-stranded chromatids for determination of chromosomal orientation and strand direction of DNA sequences (1). Standard fluorescent in situ hybridization methods rely on nucle ...

The technique of radioactive in situ hybridization of chromosomes is increasingly being replaced by nonisotopic methods. Amongst the principal reasons for this are greater convenience, resolution, safety, and speed offered by the nonradioactive methods; and the computerizat ...

The development of chromosome-specific DNA α-satellite probes, chromosome paints and contigs, together with the refinement of fluorescence in situ hybridization (FISH) techniques, has added much to the power of the technique for the detection of specific chromosomes in sperm. The te ...

In situ hybridization is a technique whereby a labeled DNA probe is hybridized directly to denaturated metaphase chromosomes fixed on a microscope slide. The cloned probe, labeled by a radioactive or fluorescent tag, hybridizes specifically to the single-stranded sequence(s) in the ge ...

Since its development by Pardue and Gall (1), the technique of in situ hybridization to polytene chromosomes has played a central role in the molecular genetic analysis of Drosophila melanogaster. The power of in situ hybridization is due largely to the scale of polytene chromosomes and conse ...

In situ hybridization of nucleotide sequence to Drosophila melanogaster interphase polytene chromosomes is a well established procedure. The method was initially developed using radioactively labeled probes by Pardue, who has recently published an extensive account of Dros ...

In the 1930s, the discovery of a simple method for the isolation and detailed microscopic observation of the banded structures that lie within the nuclei of salivary gland cells of Drosophila melanogaster was soon followed by the realization that these structures were in fact a highly amplif ...

Chromosomal in situ suppression (CISS) hybridization with flow-sorted chromosome libraries (FSCL) has provided a very powerful tool in human cytogenetics. This technique allows the painting of specific chromosomes in metaphase spreads and in interphase nuclei (1–4). The useful ...

Detecting viral infections depends largely on serological methods to demonstrate the presence of viral circulating antibodies and molecular techniques to identify viral genomes in tissue extracts. Although sensitive, none of these provides the simultaneous morphological ...

Unlike other applications of in situ hybridization (ISH),where the target is likely to be chromosomal DNA or RNA transcripts, the target in studies to detect viral nucleic acids may represent double-or single-stranded (ds or ss) DNA or RNA (representative of viral genomes), nucleic acid repl ...

It has been possible for a number of years to detect RNA in situ in fixed cells or whole tissue-sections by the use of cloned probes (1–4). The probes are labeled either with radioactive nucleotides or modified nonradioactive nucleotides, such as biotinylated dUTP, which can be visualized by fluore ...

The technique of hybridization histochemistry (1) was developed as a means of locating specific mRNA populations in tissue sections using radiolabeled recombinant DNA probes with a complementary nucleotide sequence to the target mRNA. The original method used frozen sections fix ...

Quantitative in situ hybridization has emerged as a powerful technique for the study of gene expression within complex heterocellular systems, such as the central nervous system, by allowing the measurement of the relative levels of mRNA and their variations under experimental or phys ...

In basic and applied biomedical research there is a considerable interest in having reliable RNA in situ hybridization techniques, since these allow the detection of RNA expression in a morphological context at the individual cell level. Conventional molecular biology techniques f ...