描述

The PIKA Weihenstephan™ SO Detection Kit B Alicyclobacillus acidoterrestris enables the rapid and specific detection of Alicyclobacillus acidoterrestris by real-time PCR. Alicyclobacillus acidoterrestris, an acidophilic, spore-forming gram-positive bacteria that is also heat resistant is a common cause of fruit and vegetable juice spoilage. This kit, which includes reagents for both DNA sample preparation and real-time PCR-based detection, enables specific detection of Alicyclobacillus acidoterrestris in enriched juice samples usually in ~2 hours. Because analysis is performed directly on contaminated or 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 (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 juice 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 Alicyclobacillus acidoterrestris.

In the reaction mix, a TaqMan® primer/probe set targets a DNA sequence specific to Alicyclobacillus acidoterrestris. PCR amplification yields a fluorescence signal proportional to the amount of Alicyclobacillus acidoterrestris 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.

Simple-to-Follow Workflow
Juice samples are typically directly enriched or filtered and then the filter placed in enrichment broth. After the enrichment period, 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 B Alicyclobacillus acidoterrestris 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:	A. acidoterrestris
Validated Application:	Real-Time PCR
Detection Method:	Primer-Probe Detection
Platform:	7500 Fast System, StepOnePlus™ System, StepOne™ System
Sample Type (Specific):	Fruit Juice Samples, Brewery Samples, Enriched Wine Samples, Vegetable Juice Samples, Enriched Beer Samples
Product Size:	48 reactions

内容及储存