This kit provides a convenient and rapid method to purify total RNA from small amounts of stool samples. All types of stool samples can be processed with this kit, including animal fecal samples, manure and samples collected using Norgen’s Stool Nucleic Acid Collection and Transport Devices Dx (Cat. Dx45660). The kit removes all traces of humic acids using rapid and simple spin column procedures. Bead tubes are also provided for effective homogenization of stool. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA and small interfering RNA. Both host and microbial RNA is recovered. The protocol does not rely on the use of phenol or chloroform, thereby providing a user friendly procedure and allowing high-throughput analysis. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR and reverse transcription PCR for gene expression analysis. The procedure can be completed in approximately 30 minutes.
NOTE: This product is not available for sale in the United States.
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| Kit Specifications | |
| Maximum Stool Input | 200 mg |
| Type of Stool Processed | Frozen and fresh stool |
| Maximum Column Binding Capacity | 50 μg |
| Maximum Column Loading Volume | 650 μL |
| Time to Complete 10 Purifications | 30 minutes |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
| Component | Cat. Dx49500 (50 preps) |
|---|---|
| Lysis Solution | 60 mL |
| Wash Solution | 22 mL |
| Elution Buffer | 6 mL |
| Bead Tubes | 50 |
| Spin Columns | 50 |
| Collection Tubes | 50 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
Intended Use
For the selective separation and enumeration of enterococci in food and water.
Principle and Interpretation
Tryptone and peptone are the sources of nitrogen and essential growth factors. Yeast extract acts as well nitrogenous compounds and additionally the vitamin B12 complex. Sodium azide acts largely inhibits the growth of gram-negative bacteria while sparing enterococci, staphylococci and streptococci. Ox bile inhibits most gram positives but not enterococci. Enterococci hydrolyse esculin to esculetin and dextrose, which reacts with ferric citrate producing a brownish black precipitate around the colonies. Tolerance to bile and the ability to hydrolyze esculin is the traditional and reliable test for the identification of enterococci. (4). Sodium chloride maintains the osmotic balance of the medium and Agar is the solidifying agent.
Formulation
| Ingredients | /liter |
| Tryptone | 17.0g |
| Ox bile | 10.0g |
| Yeast extract | 5.0g |
| Sodium chloride | 5.0g |
| Peptone | 3.0g |
| aesculin | 1.0g |
| Ferric ammonium citrate | 0.5g |
| Sodium azide | 0.15g |
| Agar | 15.0g |
| pH 7.1±0.1 at 25°C | |
Preparation
Weigh 56.6g of dry powder of this product, add 1 L of distilled water or deionized water, stir, heat and boil until completely dissolved, and sterilize at 121℃ for 15 min.
Quality Control
Cultural characteristics observed after incubation at 35-37°C for 20-24hours.
| Quality control strains | Growth | Colony color |
| Enterococcus faecalis ATCC29212 | PR≥0.7 | Brown-black halo |
| Escherichia coli ATCC25922 | inhibited | Absence of brown-black halo |
Sorage and Shelf Life
Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.
Precautions
1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort
2. Keep container tightly closed after using to prevent clumping.
Waste Disposal
Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.
Intended Use For the selective separation and enumeration of enterococci in food and water. Principle and Interpretation Tryptone and peptone are the sources of nitrogen and essential grow……
Bacillus cereus is a rod-shaped, gram-positive bacterium. Some strains of B. cereus cause foodpoisoning and other diseases such as keratitis. B. cereus grows at a wide range of temperature, from 4°C to 37°C. In fact, many foodborne illnesses caused by B. cereus are a consequence of improperly cooked or improperly stored food, including dairy products and meats. The majority of food poisoning by B. cereus could be divided into two types of symptoms – the emetic type and the diarrheal type, both of which are caused by the enterotoxin produced by the bacteria.
Bacillus cereus TaqMan PCR Kit,
100 reactions
Bacillus cereus TaqMan PCR Probe/Primer Set and Controls,
100 reactions
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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
| Component | Cat. TM36950 (100 preps) | Cat. TM36910 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| Bacillus cereus Primer & Probe Mix | 280 μL | 280 μL |
| Bacillus cereus Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
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