MicroSeqTM Fungal LSU rRNA Gene Sequencing (PE Applied Biosystems)
DNA is extracted from fungal cells with enzymes. The LSU rRNA gene is
selected and amplified using primers and thermalcyclers. The LSU rRNA
gene product is sequenced using cycle sequencing chemistry. The
reactions are analyzed using automated DNA sequencers and software.
Unknown fungal samples are identified using the MicroSeqTM
software and LSU rRNA database, containing over 1,500 entries. Either
the entire LSU rRNA gene, or a smaller portion of the gene (300bp) can
be sequenced. A printout of the sequence data and the identification is
included in each report.
GC-FAME (Gas Chromatography Fatty Acid Methyl Ester)
Also
known as Cellular Fatty Acid Analysis, allows for microorganism
identification based on the specific fatty acid composition of the cell
wall. Fatty acids are extracted from cultured samples and are separated
by gas chromatography. A computer generated, unique profile pattern of
the extracted fatty acids (9 to 20 carbon chains long) are compared to
our microbial databases of well over 2600 species and subspecies. Our
reports include the fatty acid profiles, a listing of the best database
matches, along with an assigned statistical probability value
indicating the confidence level of the identifications.
Fungal BIOLOG
This automated, identification system uses a 96-well microtiter plate with 95 different carbon sources. The microorganism of interest is applied to each well. Each microorganism has unique capacities to oxidize some of the various carbon sources. When these carbon sources are oxidized by the microorganism, a purple dye develops visible pattern of positive (purple) and negative (clear) well which provide a metabolic signature of the organism. The system's computer examines the patter signature with its database to determine an identification. Databases include aerobic and anaerobic bacteria, yeasts, and fungi.