Different-sized amplicons from 3 ng of a DNA plasmid were amplified. The use of Taq DNA polymerase resulted in clean and high yield of products, as analysed after PCR on a 0.8% agarose gel.
A fragment (64 bp) of the human blood-coagulation factor IIa (F2) was amplified from 20 ng, 2 ng, 200 pg and 20 pg of a human genomic DNA extract. The same experiment was performed in parallel using the Taq DNA polymerase mix from another well-established and known supplier. PCR products were subsequently analysed on a 2.5% agarose gel.
Non-specific amplification is prevented by using this Taq DNA polymerase variant as the DNA polymerase is hotstart-formulated. The 10x reaction buffer supplied together with the DNA polymerase has been specifically designed for optimal PCR performance and DNA polymerase activity. This allows the use of this DNA polymerase in a wide range of PCR applications.
Listeria monocytogenes have emerged as significant foodborne pathogens that pose a serious public health problem. As the causative agent of Listeriosis, L. monocytogenes has the highest rate of fatality rate among all foodborne pathogens. L. monocytogenes is a facultatively intracellular, Gram-positive bacterium that could survive high and low temperatures, low pH. It is a rare causative agent of mastitis in cow. However, due to its ability to resist various food processing technologies as well as to grow at low temperature, L. monocytogenes is know to be associated with both raw and pasteurized milk, as well as dairy products such as cheese. As little as 1000 organisms may cause the disease with pregnant, new-born, and immunocompromised individuals the most susceptible.
Listeria monocytogenes TaqMan PCR Kit, 24 reactions
Figure 1 / 3
Click for expanded view
Storage Conditions
The Listeria monocytogenes TaqMan PCR Kit Dx is shipped on dry ice. The components of the kit should be frozen upon arrival. If one or more of the components is not frozen when the kit is received, or if any of the components have been compromised during shipment, please contact Norgen Biotek for assistance. All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 3 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. DxTM30400 (24 rxns) |
|---|---|
| MDx TaqMan 2X PCR Master Mix Dx | 550 µL |
| L. monocytogenes Primer Mix Dx | 2 x 70 µL |
| L. monocytogenes Positive Control Dx – 200,000 copies/μL | 50 µL |
| Nuclease-Free Water | 2 x 1.25 mL |
| Product Insert | 1 |
These kits provide a rapid method for the isolation and purification of total RNA and DNA sequentially from a single sample of cultured animal cells and tissues, blood, bacteria, yeast, or fungi. The lysate is passed over two columns: 1) a DNA column and 2) an RNA column. Total RNA of all sizes is purified, including microRNA. Both DNA and RNA are of the highest purity and yield.
These kits are ideal for researchers who are interested in studying the genome and transcriptome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/DNA Purification Kits are especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as they eliminate the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and DNA are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications
RNA/DNA Purification Kit (Spin Column)
Maximum column binding capacity of 50 μg for RNA and 20 μg for DNA.
RNA/DNA Purification Micro Kit (Micro)
The purified RNA and DNA fractions can be eluted in as little as 20 μL. Ideal for cell number inputs of 500,000 and as little as 5 cells. Maximum column binding capacity of 35 μg for RNA and 10 μg for DNA.
Figure 1 / 4
Click for expanded view
| Kit Specifications | |
| Maximum Column Binding Capacity | 50 μg for RNA 20 μg for DNA |
| Maximum Column Loading Volume | 650 μL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Maximum Amount of Starting Material: Animal Cells Animal Tissues Blood Bacteria Yeast Fungi Plant Tissues | 5 x 106 cells 25 mg (for most tissues)** 200 μL 1 x 109 cells 1 x 108 cells 50 mg 50 mg |
| Time to Complete 10 Purifications | 30 minutes |
| Average Yield*: HEK 293 Cells (1 x 106 cells) HEK 293 Cells (1 x 106 cells) Liver (15 mg) Liver (15 mg) | 10-15 μg RNA 2-4 μg DNA 30-35 μg RNA 4-6 μg DNA |
*Average Yield will vary depending upon a number of factors including species, growth conditions used, and development stage.
**Tissue inputs of up to 40 mg may be used, however for inputs greater than the recommended 25 mg, cross-contamination of the RNA and DNA fractions is possible.
Storage Conditions and Product Stability
Store Proteinase K at -20°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 48700 (50 preps) | Cat. 50300 (50 preps) |
|---|---|---|
| Buffer SKP | 40 mL | 40 mL |
| Wash Solution A | 2 x 38 mL 1 x 18 mL | 2 x 38 mL 1 x 18 mL |
| Elution Solution A | 6 mL | 6 mL |
| Elution Buffer F | 15 mL | 6 mL |
| RNase-Free Water | 40 mL | 40 mL |
| Proteinase K | 2 x 12 mg | 2 x 12 mg |
| gDNA Purification Columns | 50 | – |
| gDNA Purification Micro Columns | – | 50 |
| RNA Purification Columns | 50 | – |
| RNA Purification Micro Columns | – | 50 |
| Collection Tubes | 100 | 100 |
| Elution Tubes (1.7 mL) | 100 | 100 |
| Product Insert | 1 | 1 |