Sensitivity enhancement of fluorescence in situ hybridization on plant chromosomes.
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.
Busch
W
W
Botanisches Institut, Ludwig-Maximilians-Universität, München, Germany.
Martin
R
R
Herrmann
R G
RG
D002875:Chromosomes / Q000648:ultrastructure*; D001467:Hordeum / Q000648:ultrastructure; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007231:Infant, Newborn; D010944:Plants / Q000235:genetics*; D012680:Sensitivity and Specificity; D014908:Triticum / Q000648:ultrastructure