Premium
Isolation of total RNA from dermatophytes
Author(s) -
Rezaie S.,
Pourmojib M.,
Tschachler E.
Publication year - 1999
Publication title -
mycoses
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.13
H-Index - 69
eISSN - 1439-0507
pISSN - 0933-7407
DOI - 10.1046/j.1439-0507.1999.00526.x
Subject(s) - dermatophyte , rna , trichophyton , ribosomal rna , mycelium , rna extraction , trichophyton rubrum , microbiology and biotechnology , biology , northern blot , proteinase k , microsporum canis , chemistry , biochemistry , botany , dna , gene , antifungal
We report a method for the preparation of total RNA from the anthropophilic dermatophyte Trichophyton rubrum . To generate large quantities of mycelia, the fungus was grown in liquid culture medium. The harvested mycelial mass was ground to a fine powder in liquid nitrogen and homogenized in guanidine isothiocyanate buffer followed by ultracentrifugation of the obtained suspension through a caesium chloride gradient. Analysis of the prepared RNA showed two prominent ribosomal RNA (rRNA) bands of about 3.36 and 1.82 kb. Northern blot hybridization with a β‐actin cDNA confirmed the high quality of the fungal mRNA. Successful isolation of RNA from two other dermatophyte species, namely Trichophyton mentagrophytes and Microsporum canis , demonstrated the general applicability of the described procedure.