Taq Mammalian Tissue PCR Kit
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Price:
US$522.00
(Size: 100 rxns)
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Abbexa's Taq Mammalian Tissue PCR Kit is a fast and efficient solution for DNA extraction from a variety of mammalian tissues. DNA extraction is performed in a single tube without the need for multiple washing steps, reducing the risk of sample loss and contamination.
Contents: | Component | 100 U | 500 U |
| Buffer A | 2 × 1 ml | 10 × 1 ml |
| Buffer B | 1 ml | 5 × 1 ml |
| Taq HS Red Mix | 1.25 ml | 5 × 1.25 ml |
| Target |
Mammalian Tissue |
| Tested Applications |
PCR |
| Form |
Liquid |
| Storage |
Store at -20 °C. Avoid repeated freeze/thaw cycles. |
| Validity |
Up to 12 months. |
| Buffer |
The exact formulation is proprietary. |
| Concentration |
2X |
| Availability |
Shipped within 3-7 working days. |
| Note |
This product is for research use only. Not for human consumption, cosmetic, therapeutic or diagnostic use. |
| Directions for use |
Reaction Components: | Component | Volume | | Template | 1-2 µl | | Primers (20 µM each) | 0.5 µl | | Taq HS Red Mix (2X) | 12.5 µl | | Water | Variable, up to 25 µl | | Total Volume | 50 µl | Thermal Cycling Conditions: | Step | Number of Cycles | Temperature | Time per Cycle | | Initial Denaturation | 1 cycle | 95 °C | 3 min | | Denaturation | 35 cycles | 95 °C | 15 seconds | | Annealing | ≥ 55 °C
(primer dependent) | 15 seconds | | Extension | 72 °C | 20 seconds | Notes: - Forward and reverse primers are generally used at a final concentration of 0.2-0.6 µM each. It is recommended to start with 0.4 µM as the final concentration (i.e. 10 pmol of each primer per 25 µl reaction volume). A primer concentration that is too high can reduce the specificity of priming, resulting in non-specific products. Primers should have a melting temperature (Tm) of approximately 60 °C.
- The initial denaturation step at 95 °C for 3 minutes is required to activate the enzyme and fully melt the template.
- It is recommended to run the subsequent denaturation steps at 95 °C for 15 seconds each cycle, which is suitable for GC-rich templates.
- The optimal annealing temperature is primer dependent and is usually 2-5 °C below the lower Tm of the pair. It is recommended to start with an annealing temperature of 55 °C and, if necessary, run a temperature gradient to determine the optimal annealing temperature. 15 seconds per cycle is usually sufficient, though increasing this step up to 45 seconds may improve problematic reactions.
- The optimal extension time is dependent on the length of the amplicon and the complexity of the template. An extension time of 15 seconds is sufficient for amplicons under 1 kb. Longer extension times are recommended for fragments larger than 1 kb. The extension time may be increased up to 45 seconds/kb to find the fastest optimal condition.
- The optimal conditions will vary from reaction to reaction and are dependent on the system used. Each parameter needs to be adjusted by the end user and some optimization may be required.
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Research Articles on Mammalian Tissue
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