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Detection of specific RNA sequences in yeast by in situ colony hybridization
Author(s) -
Inanov Ivan,
Mironova Rumjana,
Philipova Doranelly,
Venkov Pencho
Publication year - 1990
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320060103
Subject(s) - biology , yeast , rna , oligonucleotide , microbiology and biotechnology , in situ hybridization , dna , nucleic acid thermodynamics , dna–dna hybridization , gene , biochemistry , messenger rna
Recently a convenient method for detection of specific RNA sequences in bacteria has been developed (Ivanov and Gigova, 1986) 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 × SSC, 10% formaldehyde at 65°C for 30 min; (c) hybridization with 32 P‐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. colsi colonies.