title: Detection of specific RNA sequences in yeast by in situ colony hybridization.
abstract: Recently a convenient method for detection of specific RNA sequences in bacteria has been developed but the original protocol was inapplicable to microorganisms with a rigid cell wall. Here we report a modification of the RNA colony hybridization for use with yeast. The modified method includes the following consecutive procedures: a) treatment of the yeast colonies on the membrane filter with 10% SDS at 65% C for 30 min; b) treatment of the same filter with 3 x SSC, 10% formaldehyde at 65 degrees C for 30 min; c) hybridization with 32P-labelled oligonucleotide (or DNA) specific for the RNA sequence of interest. The intensity of the radioactive signals thus obtained is comparable with that of the E. coli colonies.
authors.affiliation: Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia.
authors.forename: I
authors.initials: I
authors.lastname: Ivanov
authors.forename: R
authors.initials: R
authors.lastname: Mironova
authors.forename: D
authors.initials: D
authors.lastname: Philipova
authors.forename: P
authors.initials: P
authors.lastname: Venkov
mesh_terms: D002473:Cell Wall / Q000378:metabolism; D006868:Hydrolysis; D009693:Nucleic Acid Hybridization; D012331:RNA, Fungal / Q000032:analysis*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D012331:RNA, Fungal