PCRmax Dekkera bruxellensis Instruction Manual

26S ribosomal RNA

Dekkera bruxellensis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Dekkera bruxellensis (also known by its anamorph ‘Brettanomyces bruxellensis’) is a yeast
native to the Senne valley near Brussels, Belgium. It is one of several members of the
genus Brettanomyces in the wild, it lives on the skins of fruit.

Dekkera bruxellensis is a hemiascomycetes yeast with a genome size ranging from under
20 to over 30 Mb. It has a high ethanol tolerance, in the wine industry it does not generally
start growing until completion of the primary Saccharomyces fermentation. It is resistant to
inhibitors formed during pretreatment of lignocellulose feed stock and it is able to utilize a
number of sugars created during processing which are not used by Saccharomyces:
cellibiose and arabinose.

The yeast D. bruxellensis can cause enormous economic losses in wine industry due to
production of phenolic off-flavor compounds. D. bruxellensis is a distant relative of baker's
yeast Saccharomyces cerevisiae. Nevertheless, these two yeasts are often found in the
same habitats and share several food-related traits, such as production of high ethanol
levels and ability to grow without oxygen. In some food products, like lambic beer, D.
bruxellensis can importantly contribute to flavor development. D. bruxellensis has several
independently duplicated ADH and ADH-like genes, which are likely responsible for
metabolism of alcohols, including ethanol, and also a range of aromatic compounds.

D. bruxellensis plays a key role in the spontaneous fermentation of typical Belgian beer
styles such as Lambic, Flanders red ales, Gueuze, Kriek, and Orval. It competes with
brewer's yeast, as well as other microorganisms, in fermenting the wort, and gives the beer
a distinctive taste, which is a desired part of the style. In the wine industry, B. bruxellensis
is generally considered a spoilage its metabolic products can impart undesirable aromas to
wine.
It occurs more commonly in some vineyards than others, so producers can avoid
purchasing grapes from such sources. Used wine barrels purchased from other vintners
are another common source.

Introduction to Dekkera bruxellensis

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MIN

MAX

The PCR Max qPCR Kit for Dekkera bruxellensis (D.bruxellensis) genomes is designed for
the in vitro quantification of D.bruxellensis genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the D.bruxellensis genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
D.bruxellensis sequences based on a comprehensive bioinformatics analysis.

If you require further information, or have a specific question about the detection profile of
this kit then please send an e.mail to [e-mail] and our bioinformatics team will
answer your question.

Specificity

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Kit Contents

• D.bruxellensis specific primer/probe mix (150 reactions BROWN)

FAM labelled

• D.bruxellensis positive control template (for Standard curve RED)

• Internal extraction control primer/probe mix (150 reactions BROWN)

VIC labelled as standard

• Internal extraction control DNA (150 reactions BLUE)

• Endogenous control primer/probe mix (150 reactions BROWN)

FAM labelled

• RNAse/DNAse free water (WHITE)

for resuspension of primer/probe mixes and internal extraction control DNA

• Template preparation buffer (YELLOW)

for resuspension of positive control template and standard curve preparation

Reagents and equipment to be supplied by the user

Real-Time PCR Instrument

DNA extraction kit

This kit designed to work well with all processes that yield high quality DNA with minimal
PCR inhibitors.

Lyophilised 2x qPCR Mastermix

This kit is designed to be compatible with all commercially available Mastermixes that run
with standard cycling conditions.

Pipettors and Tips

Vortex and centrifuge

Thin walled 1.5 ml PCR reaction tubes

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