In situ hybridization to RNA in plant tissue

1987 ◽  
Vol 5 (1) ◽  
pp. 242-250 ◽  
Author(s):  
Elliot M. Meyerowitz
Keyword(s):  
In Situ Hybridization ◽  
Plant Tissue

scholarly journals In situ hybridization for the detection of rust fungi in paraffin embedded plant tissue sections

Plant Methods ◽  
2016 ◽  
Vol 12 (1) ◽  
Author(s):  
Mitchell A. Ellison ◽  
Michael B. McMahon ◽  
Morris R. Bonde ◽  
Cristi L. Palmer ◽  
Douglas G. Luster
Keyword(s):  
In Situ Hybridization ◽  
Plant Tissue ◽  
Rust Fungi ◽  
Tissue Sections

Clear visualization of the products of nonradioactive in situ hybridization in plant tissue by simple dark-field microscopy

Micron ◽  
1997 ◽  
Vol 28 (3) ◽  
pp. 185-187 ◽  
Author(s):  
Sachihiro Matsunaga ◽  
Shigeyuki Kawano ◽  
Tetsuya Higashiyama ◽  
Noriko Inada ◽  
Tsuneyoshi Kuroiwa
Keyword(s):  
In Situ Hybridization ◽  
Plant Tissue ◽  
Dark Field ◽  
Dark Field Microscopy ◽  
Nonradioactive In Situ Hybridization

scholarly journals Optimization of RNA In Situ Hybridization for mRNA Localization Detection in Mature Tissue of Cucumber Seedlings

Plants ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 1461
Author(s):  
Zixi Liu ◽  
Xi Hu ◽  
Jing Nie ◽  
Xiaojun Li ◽  
Qing Wang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Gene Function ◽  
Plant Tissue ◽  
Mrna Localization ◽  
Gene Probe ◽  
Hybridization Technique ◽  
Transformation Rate ◽  
Cucumber Seedlings ◽  
Rna In Situ Hybridization

Cucumber (Cucumis sativus L.) is one of the main vegetable crops in China. The physiological cultivation mechanism and gene function characteristics of cucumber are of great significance to the construction of modern agriculture. Due to the low genetic transformation rate of cucumber, only in situ hybridization, which does not involve the progress of gene modified transformation, is convenient to study mRNA localization, so it is more suitable for determination on mRNA localization in the mature tissue of cucumber. At present, the existing in situ hybridization technology system is more suitable for cucumber meristem than for the mature tissue of cucumber seedlings. Therefore, we optimized the traditional plant in situ hybridization protocol. Taking a known gene CsNPF7.2 (Nitrate Transporter Families protein) as an example, we then optimized the steps of plant tissue culture, gene probe preparation, plant material sampling and fixation, preparation of cross section, hybridization pretreatment, hybridization incubation, chromogenic reaction, microscopy examination, and treatment after reaction termination in order to obtain a new RNA in situ hybridization technique suitable for identification on mRNA localization in mature tissues of cucumber seedlings. This optimized technique will ensure the yield of probes, the integrity of RNA molecules, and the clarity and integrity of plant tissue structure, which is conducive to the study of gene function and screening of key genes in cucumber.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

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