Invitrogen™ Platinum™ SuperFi II DNA Polymerase

Product Code.	No. of Reactions	unitSize
16410771	100 Reactions	100 reactions
16552642	500 Reactions	500 reactions
17112949	5 x 500 Reactions	Pack of 5

Product Code. 16410771 Supplier Invitrogen™ Supplier No. 12361010

Description

Invitrogen Platinum SuperFi II DNA Polymerase is a proofreading DNA polymerase that combines superior fidelity with an innovative buffer, enabling universal primer annealing for the highest success in PCR.

Invitrogen Platinum SuperFi II DNA Polymerase is a proofreading DNA polymerase that combines superior fidelity with an innovative buffer, enabling universal primer annealing for the highest success in PCR. Due to the unique composition of the SuperFi II PCR buffer, the annealing temperature is 60°C for most primer pairs designed following the general design rules.

Features of Platinum SuperFi II DNA Polymerase include:

• Exceptional >300X Taq fidelity
• Universal primer annealing at 60°C
• Superior specificity, sensitivity, and yields
• Robust amplification of difficult-to-amplify targets

Platinum SuperFi II DNA Polymerase is an engineered enzyme with high processivity and increased resistance to PCR inhibitors. It also enables fast-cycling protocols and amplification of long targets (up to 20 kb). Platinum hot-start technology is based on proprietary antibodies that inhibit enzyme activity until the initial PCR denaturation step, preventing non-specific amplification and primer degradation. This technology also enables reaction setup at room temperature and provides increased sensitivity and yield.

Isostabilizing molecules in the buffer increase primer-template duplex stability during the annealing step and contribute to enhanced specificity without the need to optimize the annealing temperature for each primer pair. With Platinum SuperFi II DNA Polymerase, 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.

Applications

• High-fidelity PCR
• Cloning and sub-cloning
• Site-directed mutagenesis
• Amplification of GC-rich templates
• Template generation for sequencing
• High-throughput PCR
• Amplification of samples with suboptimal purity
• Long PCR

Specifications

• Content And Storage: • Platinum SuperFi II DNA Polymerase (100 μL)
  • 5X SuperFi II Buffer (1.25 mL)
  
  Store at –20°C.
• GC-Rich PCR Performance: High
• Polymerase: Platinum SuperFi II DNA Polymerase
• Reaction Speed: Fast
• Quantity: 100 reactions
• Shipping Condition: Dry Ice
• For Use With (Application): Hot-start PCR, High-fidelity PCR
• Fidelity (vs. Taq): >300X
• Hot Start: Built-In Hot Start
• No. of Reactions: 100 Reactions
• Overhang: Blunt
• Reaction Format: Standalone
• Size (Final Product): 20 kb or less

Frequently Asked Questions (FAQs)

What is the longest amplicon I can get using Platinum SuperFi II DNA Polymerase?

Platinum SuperFi II DNA Polymerase enables amplification of long targets (up to 20 kb).

I'm having a hard time amplifying/cloning low copy cDNAs via PCR. Do you have any suggestions for me?

You may want to consider using our Platinum SuperFi II DNA Polymerase (Cat. No. 12361010) and try optimizing your PCR conditions if needed. The Platinum SuperFi II DNA Polymerase demonstrates a superior detection sensitivity with as little as 0.4 ng of genomic DNA. It is also a good idea to include a positive control reaction in parallel to your test PCR to ensure there are no issues with your reagents during handling or use over time. Please note that control templates may be ordered through our GeneArt Gene Synthesis (de novo sequence with 100% guarantee delivered in a vector) or Strings (up to 3 Kb, blunt-ended dsDNA pools, cloned into ZeroBlunt TOPO vector, convenient for multiple controls) services.

What are your recommendations when working with long amplicons using Platinum SuperFi II DNA Polymerase?

Good lab practices are important for long fragment amplification. These include using high-quality templates (pure, fresh, and intact) and fresh primer solutions. Reducing primer concentration to 0.2 µM may also improve the results.

Can I use Platinum SuperFi DNA Polymerase cycling protocol and primer annealing temperature with Platinum SuperFi II DNA Polymerase?

Yes, Platinum SuperFi II DNA Polymerase works at both universal 60 degrees C annealing temperature and at annealing temperatures calculated with a Tm calculator.

Can I perform TA cloning with Platinum SuperFi II DNA Polymerase?

Platinum SuperFi II DNA Polymerase produces blunt-ended PCR products that can be cloned directly into blunt-ended cloning vectors. TA cloning is also possible if 3' dA-overhangs are added after PCR. We recommend purifying the PCR products of Platinum SuperFi DNA Polymerase before adding the overhangs. The procedure for adding 3' dA-overhangs (TA cloning) includes the following steps:
- Purify the PCR product (e.g., with a PCR purification kit or phenol extraction/DNA precipitation). Before adding the overhangs, PCR product must be purified, as the strong proofreading activity of any remaining Platinum SuperFi DNA Polymerase will degrade the added 3' dA-overhangs.
- Perform 3' dA addition with a Taq DNA polymerase.
Reaction components:
Purified PCR product
0.2 mM dATP
1x Taq Buffer
1 U Taq DNA Polymerase
Incubate the reaction for 20 min at 72 degrees C.
- Proceed to TA cloning. For optimal ligation efficiency, we recommend using fresh PCR products, since 3' dA-overhangs can be lost during storage

Can I perform restriction digestion in the Platinum SuperFi II reaction buffer?

Platinum SuperFi II reaction buffer is compatible with restriction digestion directly after PCR. Note that Platinum SuperFi II DNA Polymerase is not inactivated during PCR, thus purification of PCR products before restriction digestion is recommended if intact 5'- or 3'- overhangs are needed for cloning.

Does the green tracking dye in Platinum SuperFi II Green PCR Master Mix interfere with downstream applications?

The green tracking dye (which consists of a blue and a yellow dye) is compatible with downstream applications such as fluorescent automated DNA sequencing, ligation, and restriction digestion. For applications that require analysis of PCR products by absorbance or fluorescence excitation, we recommend using colorless versions of the products (Platinum SuperFi II DNA Polymerase or Platinum SuperFi II PCR Master Mix) or purifying the PCR products prior to analysis.

Will the green tracking dye in Platinum SuperFi II Green PCR Master Mix interfere with fragment analysis in E-Gel agarose gels?

The green tracking dye (which consists of a blue and a yellow dye) does not interfere with electrophoresis in E-Gel agarose gels. The sample should be diluted 2- to 20-fold for optimal separation using E-Gel agarose gels.

Which nucleotide analogues can I use with Platinum SuperFi II DNA Polymerase?

Platinum SuperFi II DNA Polymerase cannot read dUTP-derivatives or dITP in DNA templates, so the use of these analogues is not recommended. Platinum SuperFi II DNA Polymerase can incorporate 7-deaza-dGTP and radiolabeled dNTPs.

Can I amplify AT-rich targets with Platinum SuperFi II DNA Polymerase?

Platinum SuperFi II DNA Polymerase enables amplification of both AT-rich and GC-rich targets. Lower extension temperature (68 degrees C or even 65 degrees C) can be helpful for extremely AT-rich targets.

Can I amplify GC-rich targets with Platinum SuperFi II DNA Polymerase?

Platinum SuperFi II DNA Polymerase can amplify targets with high GC content (up to 75% GC) without any additional DNA melting agents. In cases of extremely GC-rich targets (>75% GC), addition of DMSO to final concentration of 5% is recommended.