The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples.
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA and miRNA from cell and tissue without MagZol reagent |
| Applications | RT-PCR, Northern Blot, poly A+purification, nucleic acid protection and in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Cell, Animal tissue, Plant tissue |
| Sample amount | Cells: ≤ 5 x 10^6, Animal tissue:<10mg |
| Elution volume | ≥30μl |
| Time per run | ≤40 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
Biological samples are first lysed and homogenized in a highly denaturing guanidine isothiocyanate-containing buffer, which immediately inactivates DNases and RNases to ensure isolation of intact DNA and RNA. The lysate is then passed through a Mini spin column. This column, in combination with the high-salt buffer, allows selective and efficient binding of genomic DNA. Flow-through from the column is digested by Proteinase K in the presence of ethanol. This optimized digestion, together with the subsequent addition of further ethanol, allows appropriate binding of total RNA, including miRNA, to the column. Contaminants are efficiently washed away and high-quality RNA is eluted.
Advantages
Kit Contents
| Contents | R431102 | R431103 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Mini Columns | 100 | 2 x 250 |
| 2ml Collection Tubes | 100 | 2 x 250 |
| Proteinase K | 48 mg | 240 mg |
| Protease Dissolve Buffer | 5 ml | 15 ml |
| Buffer RLC | 40 ml | 200 ml |
| Buffer RWC | 20 ml | 80 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 60 ml |
Storage and Stability
Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples.
Toxoplasma gondii is a single-celled organism which causes Toxoplasmosis, the most common parasitic infection worldwide affecting animals as well as humans. Cats are the only animal in which sexual reproduction of the organism occurs, and for this reason cats are the only domestic animal which has the potential to shed T. gondii eggs. When disease does occur, it may develop following primary infection due to an inadequate immune response to stop the invasive tachyzoites or as a result of a reactivated infection due to a compromised immune system. The most common symptoms of the disease are anorexia, weight loss, lethargy, dyspnea, ocular symptoms and pyrexia. As toxoplasmosis can be transmitted to humans it represents a serious health risk for people living in close contact with infected animals. Infection is especially dangerous for people with suppressed immune systems and cancer patients as well as pregnant women.
Toxoplasma gondii TaqMan PCR Kit,
100 reactions
Toxoplasma gondii TaqMan PCR Probe/Primer Set and Controls,
100 reactions
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Storage Conditions
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 | TaqMan Probe Cat. TM44750 (100 preps) | TaqMan Probe Cat. TM44710 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 µL | – |
| Toxoplasma gondii Primer Probe Mix | 280 µL | 280 µL |
| Toxoplasma gondii Positive Control | 150 µL | 150 µL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
The Opentrons Flat Bottom Aluminum Block can be placed directly on the OT-2/Opentrons Flex deck or on the Opentrons Temperature Module. This block can hold flat bottom plates and other flat labware at constant temperature. The aluminum block holds temperatures between 4°C and 95°C when used with the Opentrons Temperature Module (GEN2).
The Opentrons Flat Bottom Aluminum Block can be placed directly on the OT-2/Opentrons Flex deck or on the Opentrons Temperature Module. This block can hold flat bottom plates and other flat labware at constant temperature. The aluminum block holds temperatures between 4°C and 95°C when used with the Opentrons Temperature Module (GEN2).