In situ hybridization using biotin-labeled oligonucleotides: probe labeling and procedures for mRNA detection.
نویسنده
چکیده
Methods for non-radioactive in situ hybridization are rapidly progressing in refinement and application. In this overview, we present our experiences with the use of synthetic oligodeoxynucleotides, labeled with biotin- or digoxigenin-tagged nucleotides by the terminal transferase method.
منابع مشابه
دورگه‌سازی در محل؛ اصول و کاربردها : مقاله مروری
In situ hybridization (ISH) is a method that uses labeled complementary single strand DNA or RNA to localize specific DNA or RNA sequences in an intact cell or in a fixed tissue section. The main steps of ISH consist of: probe selection, tissue or sample preparation, pre-hybridization treatment, hybridization and washing, detection and control procedure. Probe selection is one of the important ...
متن کاملDevelopment of an Alu-PCR Amplified YAC Probe Suitable for Enumeration of Chromosome 13 on Uncultured Lymphocytes and Amniocytes by Fluorescence in situ Hybridization
The main objective of the present study was to develop an efficient and reliable probe to be routinely used for detection of chromosome 13 copy numbers by interphase FISH. To achieve this, a Yeast Artificial Chromosome (YAC) containing sequences specific for human 13q12 (744D11), was cultured and the whole yeast genomic DNA was extracted. The human insert within the isolated DNA was amplified b...
متن کاملStellarisTM fluorescence in situ hybridization (FISH) probes: a powerful tool for mRNA detection
Sensitive enough to detect individual molecules of mRNA, Stellaris FISH probe sets consist of multiple singly labeled oligonucleotides (generally 30–48) designed to hybridize along targeted RNA transcripts. Each discrete transcript is observed as a diffraction-limited spot by conventional fluorescence microscopy. The relatively large number of probes used to detect a single mRNA molecule makes ...
متن کاملFastTag Nucleic Acid Labeling System: a versatile method for incorporating haptens, fluorochromes and affinity ligands into DNA, RNA and oligonucleotides.
The FastTag Nucleic Acid Labeling System couples haptens, fluorochromes or affinity ligands to any nucleic acid by attaching a universal, photo-or heat-activatable moiety to which any sulfhydryl-reactive compound can be linked. To demonstrate the versatility of the FastTag system, we have labeled DNA, RNA and oligonucleotide probes with a variety of maleimide-coupled moieties and have used thes...
متن کاملDetection of transposable elements in Drosophila salivary gland polytene chromosomes by in situ hybridization.
In situ hybridization is particularly appropriate for mapping specific DNA sequences on polytene chromosomes of Drosophila and other dipterans. This technique is based on the recognition and binding of one labeled sequence (the probe) to homologous sequences on chromosomes fixed on a microscope slide. The probes are labeled with biotin or other nonradioactive products, and the probe signal can ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Archives of histology and cytology
دوره 52 Suppl شماره
صفحات -
تاریخ انتشار 1989