ISSN:
0749-503X
Keywords:
Yeast
;
transcription
;
recombinant DNA
;
in situ hybridization
;
Life and Medical Sciences
;
Genetics
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
Notes:
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.
Additional Material:
2 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/yea.320060103
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