Invitrogen™ Platinum™ II Hot-Start PCR Master Mixes (2X)

Catalog No.	No. of Reactions	Color
14-000-012	50 reactions	Colorless
14001012	50 reactions	Green
14001013	200 Reactions	Green
14001014	1000 Reactions	Green
14-000-014	1000 Reactions	Colorless
14000013	200 Reactions	Colorless

Catalog No. 14-000-012 Supplier Invitrogen™ Supplier No. 14000012

Description

Includes

1.25 mL Platinum II PCR Master Mix (2X), 1.25 mL Platinum GC Enhancer, 1.25 mL Water, nuclease-free

Invitrogen Platinum II Hot-Start PCR Master Mix (2X) offers Platinum II Taq Hot-Start DNA Polymerase premixed with Platinum II PCR buffer and dNTPs for convenient PCR setup.

Invitrogen Platinum II Hot-Start PCR Master Mix (2X), available in colorless and green formats, offers Platinum II Taq Hot-Start DNA Polymerase premixed with Platinum II PCR buffer and dNTPs for convenient PCR setup. Platinum II Taq Hot-Start DNA Polymerase is designed for universal primer annealing and fast, easy PCR with its unique combination of innovative buffer, high-performance engineered Taq DNA polymerase and leading hot-start technology. This master mix is provided with the optional Platinum GC Enhancer for specific amplification and improved yields of GC-rich targets.

Platinum II Hot-Start Green PCR Master Mix also includes two tracking dyes for direct loading of PCR products on gels.

Features

• Innovative buffer enables universal annealing temperature by isostabilizing primer-template duplex structures
• Engineered Taq DNA polymerase confers fast cycling and resistance to common inhibitors
• Hot-start technology enables superior specificity, sensitivity, and yields and allows for room temperature reaction setup
• Green PCR buffer reduces pipetting errors with direct gel loading

Applications

• Amplification of DNA from complex genomic, viral, and plasmid templates
• Amplification and improved yields of GC-rich targets
• RT-PCR
• Genotyping
• High-throughput PCR

Platinum II Taq Hot-Start DNA Polymerase is an engineered Taq DNA polymerase that shows increased resistance to reaction inhibitors originating from sample material or DNA purification steps. The polymerase has a higher DNA synthesis rate and delivers PCR results more than two times faster than other Taq DNA polymerases. Proprietary Platinum Taq antibodies block polymerase activity at ambient temperatures and dissociate after the initial denaturation step at 94°C. 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 II PCR buffer, the annealing temperature is 60°C for most primer pairs designed following 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. With Platinum II Hot-Start PCR Master Mix (2X), different PCR assays can be cycled together using the same protocol with universal primer annealing temperature and the extension step selected for the longest fragment to be amplified.

Notes

• For applications that require PCR product analysis by absorbance or fluorescence excitation, colorless format is recommended.

Specifications

• Concentration: 2X
• Content And Storage: • Platinum II PCR Master Mix (2X), 1.25 mL
  • Platinum GC Enhancer, 1.25 mL
  • Nuclease-free water, 1.25 mL
  
  Store at -20°C in a non-frost-free freezer.
• Format: Tube
• GC-Rich PCR Performance: High
• Polymerase: Platinum II Taq Hot-Start DNA Polymerase
• Reaction Speed: Fast or Standard
• Product Type: Hot Start PCR Master Mix
• Purity or Quality Grade: HPLC
• Quantity: 50 reactions
• Shipping Condition: Dry Ice
• For Use With (Application): Hot-start PCR
• Fidelity (vs. Taq): 1X
• Hot Start: Built-In Hot Start
• No. of Reactions: 50 reactions
• Overhang: 3'-A
• Reaction Format: SuperMix or Master Mix
• Size (Final Product): 5 kb or less
• Starting Material: DNA
• Color: Colorless

Frequently Asked Questions (FAQs)

What is the storage temperature of Platinum II Taq Hot-Start DNA Polymerase products?

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.

Is the PCR product from reactions performed with Platinum II Hot-Start PCR Master Mix and Platinum II Hot-Start Green PCR Master Mix compatible with E-Gel agarose gels?

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.

Do the dyes in Platinum II Green PCR Buffer interfere with PCR performance of Platinum II Taq Hot-Start DNA Polymerase?

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.

Can Platinum II Taq Hot-Start DNA Polymerase be used in master mixes for qPCR?

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.

Can I keep my PCR reactions with Platinum II Taq Hot-Start DNA Polymerase at room temperature before the cycling starts?

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.

Can I use the same cycling protocol that I use with standard hot-start Taq for PCR reactions with Platinum II Taq Hot-Start DNA Polymerase?

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

Can I use a 2-step cycling protocol with Platinum II Taq Hot-Start DNA Polymerase, combining the annealing and extension steps?

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.

What is the DNA synthesis rate in PCR with Platinum II Taq Hot-Start DNA Polymerase?

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.

Can Platinum II Taq Hot-Start DNA Polymerase amplify GC-rich targets?

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).

My PCR with Platinum II Taq Hot-Start DNA Polymerase amplifies non-specific products. What are your recommendations?

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.

With Platinum II Taq Hot-Start DNA Polymerase and Platinum II PCR buffer, how is it possible to use an annealing temperature of 60 degrees C for any primer pair?

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.

Do my primers need to have a melting temperature (Tm) of 60 degrees C for use with Platinum II Taq Hot-Start DNA Polymerase and Platinum II PCR buffer?

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.

Safety and Handling

WARNING: Cancer - www.P65Warnings.ca.gov

For Research Use Only.