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Invitrogen™ Platinum™ Direct PCR Universal Master Mix
Description
Invitrogen Platinum Direct PCR Universal Master Mix is designed to amplify DNA directly from various samples without the need to purify DNA. It contains high-performing, engineered Platinum II Taq Hot-Start DNA Polymerase with dNTPs in an innovative buffer that enables universal primer annealing for superior performance in direct PCR applications.
Features of Platinum Direct PCR Universal Master Mix include:
- Direct DNA amplification from various samples
- Universal primer annealing at 60°C
- Optional Platinum GC Enhancer for specific amplification and improved yields of GC-rich targets
- Superior specificity, sensitivity, and yields
Platinum Direct PCR Universal Master Mix is designed to work with a variety of samples of different origins such as animal, human, plant, insect, worm, and bacterial samples. The quick lysis protocol enables efficient amplification from templates up to 8 kb in size and co-cycling of different length fragments.
Engineered Platinum II Taq DNA polymerase confers faster cycling and tolerance to reaction inhibitors originating from sample material. Proprietary Platinum Taq antibodies block polymerase activity at ambient temperatures and dissociate after the initial denaturation step. This automatic 'hot start' provides increased sensitivity, specificity, and yield, while allowing reaction assembly at room temperature.
Due to the unique composition of the Platinum Direct PCR Universal Master Mix, the annealing temperature of 60°C can be used for most primer pairs that follow the general design rules. Isostabilizing molecules in the buffer increase primer-template duplex stability during the annealing step and contribute to enhanced specificity without the need to optimize annealing temperature for each primer pair. Different PCR assays can be cycled together using the same protocol with a universal primer annealing temperature and an extension step selected for the longest fragment to be amplified.
Specifications
Specifications
| Concentration | 2X |
| Content And Storage | • 2X Platinum Direct PCR Universal Master Mix (1 mL) • Lysis Buffer (5 mL) • Proteinase K (150 μL) • Platinum GC Enhancer (1.25 mL) • Nuclease-free water (1.25 mL) Store at –20°C. |
| GC-Rich PCR Performance | High |
| Polymerase | Platinum II Taq Hot-Start DNA Polymerase |
| Reaction Speed | Fast |
| Product Type | Direct PCR Universal Master Mix |
| Quantity | 100 reactions |
| Shipping Condition | Dry Ice |
| For Use With (Application) | Hot-start PCR, Real Time PCR (qPCR) |
| Fidelity (vs. Taq) | 1X |
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Frequently Asked Questions (FAQs)
You can use any plant or tissue sample. However, when working with new sample materials, we recommend using the Lysis protocol as it allows several PCR reactions to be performed from the same sample during optimization. We also recommend having a positive control with purified sample DNA to ensure that the PCR conditions are optimal. If the positive control with purified DNA fails, the PCR conditions should be optimized before continuing further.
Proteinase K is required when PCR is performed directly from tissue samples using the Direct PCR protocol. Cell debris present in these PCR products can cause DNA fragments to get trapped in the agarose gel wells and Proteinase K helps to eliminate this.
Electrophoretic separation on E-Gel agarose gels depends on the salt concentration in the analyzed sample. For optimal band separation, we recommend diluting PCR reactions 2- to 20-fold with water, prior to running on E-Gel agarose gels. The dyes in the Platinum Direct PCR Universal Master Mix do not interfere with fragment separation on E-Gel agarose gels.
Samples can be incubated for up to 8 hr at room temperature in the Lysis Solution, prior to the 98 degrees C Proteinase K inactivation step. Please note that longer incubation may increase the product yield. For longer incubation, the Proteinase K inactivation step may be increased up to 10 min.
For the Direct protocol, the recommended sample size is up to 1 mm. For the Lysis protocol, the recommended sample size is up to 2 mm. Sample size can be increased up to 1 cm, however, Lysis Buffer volume needs to be increased to cover the whole sample.
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