Preparation of Samples for Comparative Studies of Arthropod Chromosomes: Visualization, In Situ Hybridization, and Genome Size Estimation

2005 ◽  
pp. 460-488 ◽  
Author(s):  
Rob DeSalle ◽  
T.Ryan Gregory ◽  
J.Spencer Johnston
Keyword(s):  
In Situ Hybridization ◽  
Genome Size ◽  
Comparative Studies ◽  
Size Estimation

scholarly journals Cytogenetic relationships within the Maghrebian clade of Festuca subgen. Schedonorus (Poaceae), using flow cytometry and FISH

2017 ◽  
Vol 74 (1) ◽  
pp. 052 ◽  
Author(s):  
David Ezquerro-López ◽  
David Kopecký ◽  
Luis Á. Inda
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Fluorescent In Situ Hybridization ◽  
Genome Duplication ◽  
Size Estimation ◽  
Ploidy Levels ◽  
New Information ◽  
Shed Light

Festuca subgen. Schedonorus is a group of broad-leaved fescues, which can be divided into two clades: European and Maghrebian. We employed fluorescent in situ hybridization —FISH— with probes specific for 5S and 35S ribosomal DNA and genome size estimation using flow cytometry to shed light on the determination of possible parental genomes of polyploid species of the Maghrebian clade. Our results indicate that octoploid F. arundinacea subsp. atlantigena probably originated from crossing of the tetraploids F. arundinacea subsp. fenas —2n = 4x = 28— and F. mairei —2n = 4x = 28— followed by whole genome duplication. However, a large reconstruction of karyotype and genome downsizing has been revealed. Similarly, hexaploid F. arundinacea subsp. corsica presumably resulted from the interspecific hybridization of the diploid F. pratensis and tetraploid F. arundinacea subsp. fenas. Several scenarios on the origin of decaploid F. arundinacea var. letourneuxiana are discussed. This study contributed to our knowledge on the phylogeny of broad-leaved fescues and provided new information on the karyotypes —chromosome numbers, ploidy levels and numbers and positions of rDNA loci— using FISH and genome size estimations using flow cytometry in selected taxa of this important grass genus.

Random amplified polymorphic DNA analysis, genome size, and genomic in situ hybridization of triploid viviparous onions

Genome ◽  
1999 ◽  
Vol 42 (6) ◽  
pp. 1208-1216 ◽  
Author(s):  
Jasna Puizina ◽  
Branka Javornik ◽  
Borut Bohanec ◽  
Dieter Schweizer ◽  
Jolanta Maluszynska ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genome Size ◽  
Allium Cepa ◽  
Dna Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Genomic In Situ Hybridization ◽  
Allium Species ◽  
Allium Cepa L ◽  
Genomic Dnas

Triploid viviparous onions (Allium cepa L. var. viviparum Metzg. (ALEF.), auct.), (2n = 3x = 24), are known in some countries only as a rare relic crop, while in other parts of the world they are still traditionally or even commercially cultivated. Results indicating an identical random amplified polymorphic DNA (RAPD) banding pattern and the same DNA content (2C = 43.4 pg) establish the high genetic similarity and the unique origin of the Croatian clone Ljutika and the Indian clone Pran. In order to determine the parental Allium species of these natural triploid hybrids, genomic fluorescent in situ hybridization (GISH) was applied. Biotinylated genomic DNAs from six diploid Allium species (A. cepa L., A. fistulosum L., A. roylei Stearn, A. vavilovii M. Pop. et Vved., A. galanthum Kar. et Kir., A. oschaninii O. Fedtsch.) were used as probes in this study. While probes obtained from genomic DNA of A. cepa, A. vavilovii, and A. roylei hybridized to somatic chromosomes of Ljutika probes from A. fistulosum, A. galanthum, and A. oschaninii did not. The DNA probes of A. cepa and A. roylei each completely or predominantly labelled one genome (eight chromosomes). A few chromosomes, the markers of the triploid karyotype, were not completely labelled by any probe applied. Our GISH results indicate that triploid viviparous onions might possess a complex triparental genome organization.Key words: triploid viviparous onions, Allium cepa, Allium roylei, genomic in situ hybridization, genome size, random amplified polymorphic DNA (RAPD).

scholarly journals Cytological Analyses in Melandrium album: Genome Size, C.hromosome Size and Fluorescence in situ Hybridization.

CYTOLOGIA ◽  
1994 ◽  
Vol 59 (1) ◽  
pp. 135-141 ◽  
Author(s):  
Sachihiro Matsunaga ◽  
Masahiro Hizume ◽  
Shigeyuki Kawano ◽  
Tsuneyoshi Kuroiwa
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Genome Size ◽  
Melandrium Album

Genome composition in Venezuelan spiny-rats of the genus Proechimys (Rodentia, Echimyidae). I. Genome size, C-heterochromatin and repetitive DNAs in situ hybridization patterns

1997 ◽  
Vol 78 (1) ◽  
pp. 36-43 ◽  
Author(s):  
S. Garagna ◽  
A. Pérez-Zapata ◽  
M. Zuccotti ◽  
S. Mascheretti ◽  
N. Marziliano ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genome Size ◽  
Genome Composition ◽  
Repetitive Dnas

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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