Sensitivity Enhancement of Fluorescence in Situ Hybridization on Plant Chromosomes

Overview

Journal Chromosome Res

Date 1994 Jan 1

PMID 8162315

Citations 7

Authors

W Busch

R Martin

R G Herrmann

Affiliations

Soon will be listed here.

Abstract

An improved in situ hybridization procedure is presented, based on synchronization of root meristems of barley and wheat, enzymatic digestion, a protoplast drop technique, and the use of the fluorescent dye Cy3. The combination of these approaches resulted in a significant increase of well-spread metaphases suitable for in situ hybridization as compared to squash preparations, and to a significantly enhanced number and intensity of hybridization signals as demonstrated for a B-hordein-specific low-copy probe of barley. In the case of Cy3 all metaphases displayed a signal, more than 60% of them on both chromatids of each gene-bearing chromosome.

Citing Articles

The relationship between physical and genetic distances at the Hor1 and Hor2 loci of barley estimated by two-colour fluorescent in situ hybridization.

Pedersen C, Linde-Laursen I Theor Appl Genet. 2013; 91(6-7):941-6.

PMID: 24169981 DOI: 10.1007/BF00223904.

Repeated DNA sequences isolated by microdissection. II. Comparative analysis in Hordeum vulgare and Triticum aestivum.

Busch W, Herrmann R, Hohmann U Theor Appl Genet. 2013; 93(1-2):164-71.

PMID: 24162213 DOI: 10.1007/BF00225741.

A rapid and efficient transformation protocol for the grass Brachypodium distachyon.

Christiansen P, Andersen C, Didion T, Folling M, Nielsen K Plant Cell Rep. 2004; 23(10-11):751-8.

PMID: 15503032 DOI: 10.1007/s00299-004-0889-5.

A new chromosome fluorescence banding technique combining DAPI staining with image analysis in plants.

Liu J, She C, Hu Z, Xiong Z, Liu L, Song Y Chromosoma. 2004; 113(1):16-21.

PMID: 15197560 DOI: 10.1007/s00412-004-0289-1.

Comparative genomic in situ hybridization (cGISH) analysis on plant chromosomes revealed by labelled Arabidopsis DNA.

Zoller J, Yang Y, Herrmann R, Hohmann U Chromosome Res. 2001; 9(5):357-75.

PMID: 11448038 DOI: 10.1023/a:1016767100766.

References

Gall J, Pardue M . Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci U S A. 1969; 63(2):378-83. PMC: 223575. DOI: 10.1073/pnas.63.2.378. View

Dolezel J, Cihalikova J, Lucretti S . A high-yield procedure for isolation of metaphase chromosomes from root tips of Vicia faba L. Planta. 2013; 188(1):93-8. DOI: 10.1007/BF00198944. View

Sorensen M . Mapping of the Hor2 locus in barley by pulsed field gel electrophoresis. Carlsberg Res Commun. 1989; 54(3):109-20. DOI: 10.1007/BF02908303. View

Lichter P, Tang C, Call K, Hermanson G, Evans G, Housman D . High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones. Science. 1990; 247(4938):64-9. DOI: 10.1126/science.2294592. View

JOHNSON G, Nogueira Araujo G . A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods. 1981; 43(3):349-50. DOI: 10.1016/0022-1759(81)90183-6. View

Pinkel D, Landegent J, Collins C, Fuscoe J, Segraves R, Lucas J . Fluorescence in situ hybridization with human chromosome-specific libraries: detection of trisomy 21 and translocations of chromosome 4. Proc Natl Acad Sci U S A. 1988; 85(23):9138-42. PMC: 282679. DOI: 10.1073/pnas.85.23.9138. View

Lawrence J, Villnave C, Singer R . Sensitive, high-resolution chromatin and chromosome mapping in situ: presence and orientation of two closely integrated copies of EBV in a lymphoma line. Cell. 1988; 52(1):51-61. DOI: 10.1016/0092-8674(88)90530-2. View

Schondelmaier J, Martin R, Jahoor A, Houben A, Graner A, Koop H . Microdissection and microcloning of the barley (Hordeum vulgare L.) chromosome 1HS. Theor Appl Genet. 2013; 86(5):629-36. DOI: 10.1007/BF00838719. View

Wiegant J, Galjart N, Raap A, dAzzo A . The gene encoding human protective protein (PPGB) is on chromosome 20. Genomics. 1991; 10(2):345-9. DOI: 10.1016/0888-7543(91)90318-9. View

10.

Viegas-Pequignot E, Dutrillaux B, Magdelenat H, Coppey-Moisan M . Mapping of single-copy DNA sequences on human chromosomes by in situ hybridization with biotinylated probes: enhancement of detection sensitivity by intensified-fluorescence digital-imaging microscopy. Proc Natl Acad Sci U S A. 1989; 86(2):582-6. PMC: 286516. DOI: 10.1073/pnas.86.2.582. View

11.

Wessendorf M, Brelje T . Which fluorophore is brightest? A comparison of the staining obtained using fluorescein, tetramethylrhodamine, lissamine rhodamine, Texas red, and cyanine 3.18. Histochemistry. 1992; 98(2):81-5. DOI: 10.1007/BF00716998. View

12.

Siedler H, Graner A . Construction of physical maps of the Hor1 locus of two barley cultivars by pulsed field gel electrophoresis. Mol Gen Genet. 1991; 226(1-2):177-81. DOI: 10.1007/BF00273601. View

13.

Pan W, Houben A, Schlegel R . Highly effective cell synchronization in plant roots by hydroxyurea and amiprophos-methyl or colchicine. Genome. 1993; 36(2):387-90. DOI: 10.1139/g93-053. View

14.

Graner A, Siedler H, Jahoor A, Herrmann R, Wenzel G . Assessment of the degree and the type of restriction fragment length polymorphism in barley (Hordeum vulgare). Theor Appl Genet. 2013; 80(6):826-32. DOI: 10.1007/BF00224200. View