Sensitive fluorescence in situ hybridization signal detection in maize using directly labeled probes produced by high concentration DNA polymerase nick translation.
The signal produced by fluorescence in situ hybridization (FISH) often is inconsistent among cells and sensitivity is low. Small DNA targets on the chromatin are difficult to detect. We report here an improved nick translation procedure for Texas red and Alexa Fluor 488 direct labeling of FISH probes. Brighter probes can be obtained by adding excess DNA polymerase I. Using such probes, a 30 kb yeast transgene, and the rp1, rp3 and zein multigene clusters were clearly detected.
Kato
A
A
Division of Biological Sciences, Tucker Hall, University of Missouri-Columbia, Columbia, Missouri, 65211-7400, USA.
Albert
P S
PS
Vega
J M
JM
Birchler
J A
JA
D032461:Chromosomes, Plant; D004256:DNA Polymerase I; D015342:DNA Probes / Q000138:chemical synthesis*; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation / Q000379:methods*; D015336:Molecular Probe Techniques / Q000295:instrumentation; D003313:Zea mays / Q000235:genetics*
D015342:DNA Probes; D004256:DNA Polymerase I