Open AccessPrimer-mediated in situ detection of the B-hordein gene cluster on barley chromosome 1H.
Author(s) -
Shahal Abbo,
Roy P. Dunford,
Terry E. Miller,
S. M. Reader,
Ian P. King
Publication year - 1993
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.90.24.11821
Subject(s) - biology , primer (cosmetics) , oligonucleotide , microbiology and biotechnology , in situ , hordeum vulgare , hordein , chromatin , chromosome , dna , hybridization probe , genetics , gene , computational biology , chemistry , storage protein , ecology , poaceae , organic chemistry
In situ hybridization methods allow the detection of specific DNA sequences on whole chromosomes. The technique has been widely used as a diagnostic and research tool by animal cytogeneticists, for whom detection of unique sequences on mammalian chromosomes is routinely achieved. However, detection of unique sequences on plant chromosomes is less reliable. The recently developed primer-induced in situ hybridization (PRINS) technique allows rapid and reliable in situ detection by the hybridization of primers to denatured target DNA, followed by extension with DNA polymerase in the presence of a labeled nucleotide. The use of short oligonucleotide primers could allow improved penetration of debris and highly condensed chromatin common in preparations of plant chromosomes, thus increasing the sensitivity of in situ detection. The feasibility of this approach is demonstrated by the oligonucleotide primer-mediated detection of the B-hordein gene cluster on a barley chromosome. Applications of the PRINS technique for plant cytogeneticists are discussed.