scholarly journals Direct Fluorescence In Situ Hybridization to Polytene Chromosomes

BioTechniques ◽  
1998 ◽  
Vol 24 (2) ◽  
pp. 184-185 ◽  
Author(s):  
Bozenna Zagrodzinska ◽  
Malgorzata Kloc
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Polytene Chromosomes ◽  
Direct Fluorescence

Detection of rRNA and phaseolin genes on polytene chromosomes of Phaseolus coccineus by fluorescence in situ hybridization after pepsin pretreatment

Genome ◽  
1994 ◽  
Vol 37 (6) ◽  
pp. 1018-1021 ◽  
Author(s):  
M. Nenno ◽  
K. Schumann ◽  
W. Nagl
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Ribosomal Rna ◽  
Storage Protein ◽  
Seed Storage Protein ◽  
Polytene Chromosomes ◽  
Seed Storage ◽  
Phaseolus Coccineus ◽  
Rna Genes

This is the first report of fluorescence in situ hybridization (FISH) on plant polytene chromosomes. Different protease pretreatments have been tested to improve fluorescence in situ hybridization FISH on polytene chromosomes of a plant, Phaseolus coccineus, with the aim to enable the detection of low-copy genes. The structural preservation of the chromosomes and the distinctness of the FISH signals were comparatively analysed with a probe for the ribosomal RNA genes after digestion with pepsin and trypsin. The pepsin pretreatment resulted in a general loosening of chromatin with good conservation of chromosome morphology and an increased number and density of signal points. The six nucleolus organizers exhibited significant differences in condensation. The pretreatment with pepsin enabled the detection of the low-copy genes encoding the seed storage protein phaseolin.Key words: plant, Leguminosae, ribosomal RNA genes, seed storage protein genes, protease.

scholarly journals Detection and Differentiation of Chlamydiae by Fluorescence In Situ Hybridization

2002 ◽  
Vol 68 (8) ◽  
pp. 4081-4089 ◽  
Author(s):  
Sven Poppert ◽  
Andreas Essig ◽  
Reinhard Marre ◽  
Michael Wagner ◽  
Matthias Horn
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Single Step ◽  
Detection Methods ◽  
The Novel ◽  
Antibody Staining ◽  
Uncultured Bacteria ◽  
Direct Fluorescence ◽  
Probe Set

ABSTRACT Chlamydiae are important pathogens of humans and animals but diagnosis of chlamydial infections is still hampered by inadequate detection methods. Fluorescence in situ hybridization (FISH) using rRNA-targeted oligonucleotide probes is widely used for the investigation of uncultured bacteria in complex microbial communities and has recently also been shown to be a valuable tool for the rapid detection of various bacterial pathogens in clinical specimens. Here we report on the development and evaluation of a hierarchic probe set for the specific detection and differentiation of chlamydiae, particularly C. pneumoniae, C. trachomatis, C. psittaci, and the recently described chlamydia-like bacteria comprising the novel genera Neochlamydia and Parachlamydia. The specificity of the nine newly developed probes was successfully demonstrated by in situ hybridization of experimentally infected amoebae and HeLa 229 cells, including HeLa 229 cells coinfected with C. pneumoniae and C. trachomatis. FISH reliably stained chlamydial inclusions as early as 12 h postinfection. The sensitivity of FISH was further confirmed by combination with direct fluorescence antibody staining. In contrast to previously established detection methods for chlamydiae, FISH was not susceptible to false-positive results and allows the detection of all recognized chlamydiae in one single step.

Fluorescence in situ Hybridization to the Polytene Chromosomes of Anopheles Mosquitoes

BIO-PROTOCOL ◽  
2013 ◽  
Vol 3 (16) ◽  
Author(s):  
Ai Xia ◽  
Ashley Peery ◽  
Maryam Kamali ◽  
Jiangtao Liang ◽  
Maria Sharakhova ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Polytene Chromosomes ◽  
Anopheles Mosquitoes

Direct and sensitive fluorescence in situ hybridization of 45S rDNA on tomato chromosomes

Genome ◽  
1994 ◽  
Vol 37 (6) ◽  
pp. 1062-1065 ◽  
Author(s):  
Jie Xu ◽  
E. D. Earle
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Chromosome 2 ◽  
45S Rdna ◽  
Plant Chromosomes ◽  
Signal Sensitivity ◽  
Direct Fluorescence ◽  
Hybridization Protocol ◽  
Tomato Genotypes

We describe a direct and sensitive fluorescence in situ hybridization protocol for plant chromosomes. We labelled 45S rDNA with fluorescein-12-dUTP and hybridized to somatic chromosomes of four tomato genotypes. This technique does not require posthybridization immunocytochemical amplifications. The improved signal sensitivity with this technique allowed identification of new rDNA loci on three pairs of chromosomes, in addition to the previously known locus on chromosome 2. We discuss favorable features of direct fluorescence in situ hybridization for chromosomes fixed on a slide and chromosomes or cells in suspension.Key words: direct fluorescence in situ hybridization, 45S rDNA, tomato chromosomes.

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