Platinum™ SuperFi™ DNA Polymerase, 100 units - FAQs

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11 product FAQs found

我可以使用Taq聚合酶生成我的目的基因用于定向TOPO克隆吗？

不，你的基因必须用一种校对活性聚合酶例如Platinum SuperFi DNA聚合酶或AccuPrime Pfx DNA聚合酶进行扩增以获得平末端才可用于定向TOPO克隆。

Can I use Taq polymerase to generate my gene of interest for directional TOPO cloning?

No, your gene of interest must be amplified with a proofreading polymerase such as Platinum SuperFi DNA Polymerase or AccuPrime Pfx DNA Polymerase that leaves blunt ends for directional TOPO cloning.

Which nucleotide analogues can be used with Platinum SuperFi DNA Polymerase?

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

I am having difficulties amplifying long targets using Platinum SuperFi DNA Polymerase. What are your recommendations?

Good lab practices are important for long fragment amplification. These include using high-quality templates (pure, fresh and intact) and fresh primer solutions. Optimization steps to consider include decreasing denaturation and extension temperatures, lengthening extension times as recommended in the manual, and increasing template amounts.

Can Platinum SuperFi DNA Polymerase amplify AT-rich targets?

Yes. To improve amplification of AT-rich targets, we recommend reducing the extension temperature to 68 degrees C or add 5-15 mM Tetramethylammonium Chloride (TMAC).

Can Platinum SuperFi DNA Polymerase amplify GC-rich targets?

All Platinum SuperFi product formats are supplied with 5X SuperFi GC Enhancer which is optimized to improve amplification of GC-rich targets (recommended for use with targets containing >65% GC content)

Does SuperFi Green Buffer influence performance of Platinum SuperFi DNA Polymerase?

No. The colored buffer does not interfere with PCR performance and does not change enzyme features.

Can Platinum SuperFi DNA Polymerase be used for amplification of bisulfite-converted DNA?

No. Platinum SuperFi DNA Polymerase cannot read uracil in the template strand, therefore, it is not recommended for use with bisulfite-converted DNA.

Does Platinum SuperFi DNA Polymerase add the non-template dependent 3'-A overhang?

No. Platinum SuperFi DNA Polymerase generates blunt ended products.

What is the amplicon size limit using Platinum SuperFi DNA Polymerase?

Platinum SuperFi DNA Polymerase accurately amplifies long fragments (up to 20 kb) with high yields and specificity. Amplification of even larger fragment sizes up to 40 kb has been demonstrated, but may require additional optimization of reaction conditions and primer design.

Can protocols optimized for Taq DNA Polymerase, Platinum Taq DNA Polymerase High Fidelity, AccuPrime Taq DNA Polymerase High Fidelity, Platinum Pfx DNA Polymerase, or AccuPrime Pfx DNA Polymerase be directly applied to Platinum SuperFi DNA Polymerase?

Platinum SuperFi DNA Polymerase significantly differs from many other DNA polymerases, therefore annealing rules should be adjusted by using our Tm calculator (www.thermofisher.com/tmcalculator). Due to high processivity, Platinum SuperFi DNA Polymerase also has shorter cycling times than many other DNA polymerases.