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View additional product information for Platinum™ II Hot-Start PCR Master Mixes (2X) - FAQs (14001012, 14001014, 14000013, 14001013, 14000012, 14000014)
12 product FAQs found
Platinum II Taq Hot-Start DNA Polymerase products can be stored at 4 degrees C for up to 3 months. For longer storage, we recommend storing all components at -20 degrees C.
Electrophoretic separation on E-Gel agarose gels depends on the salt concentration in the analyzed sample. For optimal separation, we recommend diluting PCR reactions performed with colorless and green Platinum II PCR Master Mixes 2- to 20-fold, prior to running on E-Gel agarose gels. The dyes in the Platinum II Hot-Start Green PCR Master Mix do not interfere with fragment separation on E-Gel agarose gels.
No. The tracing dyes (a blue and a yellow dye) in Platinum II Green PCR Buffer do not interfere with PCR performance and do not change any enzyme features.
Yes, Platinum II Taq Hot-Start DNA Polymerase can be used in qPCR master mixes for target detection and quantification in a real-time PCR instrument using probes or SYBR Green dye.
Yes. Due to stable antibody-mediated hot-start technology, Platinum II Taq Hot-Start DNA Polymerase is highly stable. The premixed reactions for PCR can be incubated at room temperature for up to 24 hr before loading in the thermal cycler, without any loss of amplification specificity.
PCR reactions with Platinum II Taq Hot-Start DNA Polymerase can be run using the same cycling protocol and annealing temperature as that for standard hot-start Taq polymerase
Yes, simple amplicons up to 1 kb with 45-65% GC sequences can be synthesized using a 2-step cycling protocol with a combined annealing/extension step at 60 degrees C. With the 2-step protocol, a denaturation step is performed for 5 sec at the increased temperature of 98 degrees C. For longer, GC-rich, and complex amplicons, or cDNA targets, we recommend using a 3-step cycling protocol.
Platinum II Taq Hot-Start DNA Polymerase contains engineered Taq DNA polymerase with increased DNA synthesis rate of 15 sec/kb at 68 degrees C extension temperature. Conveniently, the extension step can be prolonged up to 1 min/kb without a negative effect on specificity. This allows the cycling of shorter and longer amplicons together using the same protocol.
All Platinum II Taq Hot-Start DNA Polymerase product formats are supplied with Platinum GC Enhancer that is optimized to improve amplification of GC-rich targets (recommended for use with targets containing >65% GC).
Usually, primers that are well designed and work in PCR with standard Taq under standard PCR conditions, anneal specifically in Platinum II PCR buffer at 60 degrees C regardless of their Tm. If amplification of a particular template-primer pair does not give satisfactory results, we recommend redesigning the primers.
If there is no possibility for redesign, use a temperature gradient and empirically determine the optimal annealing temperature. Start with an annealing temperature that is at least 5 degrees C lower than your primer Tm.
5X Platinum II PCR Buffer contains isostabilizing molecules that increase primer-template duplex stability during the annealing step and contribute to enhanced specificity. As a result, we expect most primer pairs to anneal at 60 degrees C in this polymerase/buffer system, eliminating the need for annealing temperature optimization.
No, primers with various melting temperatures can be used. If designed following the general primer design rules, the majority of primers will anneal specifically at 60 degrees C regardless of their melting temperature.