描述

The PIKA Weihenstephan™ SO Detection Kit H Dekkera (Brettanomyces) enables the rapid and specific detection of Brettanomyces (aka "Brett") by real-time PCR. Dekkera and Brettanomyces are interchangeably used to describe a genus of non-spore forming yeast that is a common cause of wine and beer spoilage, but that at low levels produces several compounds that can yield desirable taste in wine. In beer, Dekkera may grow in the finished product causing off flavor and high pressure, leading to danger of explosion. This kit, which includes reagents for both DNA sample preparation and real-time PCR-based detection, enables specific detection of Dekkera (Brettanomyces) in enriched wine and beer samples usually in ~2 hours. Because analysis is performed directly on enriched samples, there is no need to isolate the organism to single colonies, saving time and yielding results days faster than culture-based methods.

• Fast—from enriched wine samples to results usually in ~2 hours
• Sensitive—detects as few as 1000 cells per sample, enabling detection long before visible growth
• Simple—pre-aliquotted, ready-to-use format minimizes errors and pipetting steps. Sample prep reagents included.
• Stable—shelf-stable reagents can be stored at room temperature

How the Kit Works
After enrichment of wine samples in appropriate media, such as PIKA Weihenstephan™ FastOrange™ Brett Broth* (supplied separately), and rapid DNA sample preparation using the reagents supplied in the kit, detection is accomplished by real-time PCR using the included TaqMan® assay reagents.

Because of its exquisite sensitivity and specificity, real-time PCR is an ideal method for detecting specific species of wine spoilage organisms within mixed cultures, even among high concentrations of brewer’s yeast. The spoilage organisms can often be detected from clear samples before the culture is visibly turbid, so results can be obtained days earlier than with traditional culture methods. Plating to individual colonies is unnecessary, and there is no guesswork regarding the presence of Dekkera (Brettanomyces).

In the reaction mix, a TaqMan® primer/probe set targets a DNA sequence specific to Dekkera (Brettanomyces). PCR amplification yields a fluorescence signal proportional to the amount of Dekkera (Brettanomyces) target sequence, which is detectable in the FAM™ fluorescence channel of the real-time PCR instrument. An internal positive control (IPC) is detected in the VIC® channel. The IPC is included to verify the absence of PCR inhibitors and that amplification proceeds normally in the reaction.

*We strongly recommend the use of PIKA Weihenstephan™ FastOrange™ Brett Broth to enrich beer or brewery samples prior to analysis with the PIKA Weihenstephan™ SO Detection Kit H Dekkera (Brettanomyces), since PIKA Weihenstephan™ FastOrange™ Brett Broth is optimized for the rapid and selective growth of Dekkera (Brettanomyces) and has been qualified for use with downstream PCR applications.

Simple-to-Follow Workflow
Wine or beer samples are typically enriched in an equal volume of ready-to-use PIKA Weihenstephan™ FastOrange™ Brett Broth (available separately) or filtered and the filter placed in 5 mL PIKA Weihenstephan™ FastOrange™ Brett Broth. After enrichment for 2–3 days, sample preparation is accomplished using a simple heat lysis method. Briefly, 1 mL of the enriched sample is pelleted by centrifugation, optionally washed, and then subjected to a 10-minute high temperature lysis using the included wash and lysis reagents. After a brief centrifugation step, the lysate is ready for analysis by PCR.

PCR set up is made easy with pre-aliquotted primer-probe mix. Simply add the included Rehydration Buffer, TaqMan® Universal Master Mix II-No UNG (available separately, Cat. No. 4440043), and prepared DNA sample to the supplied individual reaction tubes of dried primer-probe mix. Real-time PCR is then performed for 40 cycles. The entire process from DNA sample preparation to results takes only about two hours, with very little hands-on time.

The kit is compatible with Applied Biosystems® StepOne™, StepOnePlus™, 7500, and 7500 Fast real-time PCR systems.

Backed by Experts
PIKA Weihenstephan™ SO Detection Kit H Dekkera (Brettanomyces) was developed by and is manufactured for Life Technologies by PIKA Weihenstephan GmbH. Since 2000, PIKA Weihenstephan has been a consultant and leading provider of testing products and services for the brewing and beverage industry.

规格

Target Organism:	Dekkera spp., Brettanomyces spp.
Validated Application:	Real-Time PCR
Detection Method:	Primer-Probe Detection
Platform:	7500 Fast System, StepOnePlus™ System, StepOne™ System
Sample Type (Specific):	Enriched Beer Samples, Enriched Wine Samples, Brewery Samples
Product Size:	48 reactions

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