Use of biotin-labeled probes to map specific DNA sequences on …
A biotin-labeling technique was used to map a 120 bp rye DNA probe by in situ hybridization to somatic metaphase chromosomes of common wheat. Twenty-four hybrid- Ization sites were revealed on 11 chromosomes including 4A, 2D, 3D, 5D, and the seven B-genome chromosomes.
1) Wetzel, J.B., and A. L. Rayburn. 2000. Use of fluorescence genomic in situ hybridization (GISH) to detect the presence of alien chromatin in wheat lines differing in nuclear DNA content. Cytometry 41: 36-40. 2) Rayburn, A.L., J. Bouma and C. A. Northcott. 2001. Comparing the Clastogenic Potential of Atrazine with Caffeine using Chinese ...
The conditions for in situ hybridization and detection of the sites of hybridization are described by Rayburn & Gill (1985 a). Briefly, the chromosomes are denatured in 70% formamide in 2 x SSC at 70 °C for 10 min. The hybridization is carried out in 10% dextran sulfate at 37 °C for 6 h. A horseradish peroxidase detection system was used.
USE OF REPEATED DNA SEQUENCES AS CYTOLOGICAL MARKERS
Repetitive DNA sequences provide cytological markers necessary for studies of chromosomal identification, genome allocation, and genome orientation, and the use of biotin-labeled DNA probes allows the technique of in situ hybridization to be performed much more rapidly and with a greater degree of safety and reliability.
The in situ h2~bridization sites were detected by the appear- ance of a brown-colored precipitate over blue (Giemsa- stained) chromosomes (Lapitan et al. 1986; Rayburn and Gill
Wheat-rye translocations: Detection of chromosome breakpoints …
In situ hybridization with a blotin-labeled DNA probe was used to detect wheatrye translocations. The probe containing a 120-bp repetitive DNA sequence from rye, hybridized to the entire length of all rye chromosomes, but only to a few sites in 14 wheat chromosomes.
Historical perspective of in situ hybridization for the analysis of ...
The non isotopic in situ hybridization using biotin labeled DNA probes was first introduced in plant species by Rayburn and Gill (1985). Biotin labeled repetitive DNA sequences (120bp) of rye were used as probe to mitotic chromosomes of Triticum aestivum and double ditelosomic lines of B and D genome chromosomes and 4A of Chinese spring.
In situ hybridization is the specific annealing of a labeled nucleic acid probe to complementary sequences in fixed tissues followed by visualization of the location of the probe. The...
Use of fluorescence genomic in situ hybridization (GISH) to detect …
One parental line had previously been reported to have alien chromatin substituted for the corresponding wheat homologous chromatin. Genomic in situ hybridization was used to determine the presence or absence of rye chromatin in three cultivars and six near-isolines of wheat.
In Situ Hybridization to Plant Chromosomes | SpringerLink
2016年10月18日 · In situ hybridization is a fundamental method in modern plant molecular cytogenetics research. It cannot be overstressed that good in situ hybridization results begin with good cytological preparations. Well-spread chromosome preparations with low amounts of cytoplasm give the best hybridization signals.