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Platinum™ Taq Green Hot Start DNA Polymerase - FAQs

View additional product information for Platinum™ Taq Green Hot Start DNA Polymerase - FAQs (11966083)

4 product FAQs found

Do you have any recommendations for troubleshooting the amplification of GC-rich or problematic targets with Platinum Taq Green Hot Start DNA Polymerase?

We have the following recommendations:
- Increase the extension time from 1 min/kb to 1.5 min/kb.
- Vary total PCR cycles from 20-40; 35 cycles is typical.
- Try denaturing at 95 degrees C for 45 seconds.
- Increase the amount of template for targets >5 kb.
- Use 2.5 U of Platinum Taq for each 50 µL reaction.
- GC-rich or problematic targets work better with MgSO4 instead of MgCl2.
- For GC-rich templates, test higher annealing temperatures starting with the temperature that is equal to your primer Tm and in increments of 2 degrees C for up to six different temperatures.
- Vary the KB extender solution from 1.5 µL to 4.5 µL per 50 µL reaction.

When should the KB Extender be used that comes with the Platinum Taq Green Hot Start DNA Polymerase (Cat. Nos. 11966018, 11966026, 11966034, 11966083)?

The KB Extender enhances the amplification of GC-rich and problematic sequences, and the extension of genomic DNA targets of >5 kb. The KB Extender lowers DNA melting temperature (Tm), and the co-solvent and amplification buffer offer higher primer specificity, broader Mg concentration and annealing temperature optima, as well as improved Taq thermostabilization. Please note that primer sets that generate specific products using standard PCR buffer may not benefit from using KB Extender. Excessive use of the KB Extender may reduce yield, particularly for non-GC rich amplicons. We generally recommend varying the KB Extender solution from 1.5-4.5 µL per 50 µL reaction.

Is the green dye that comes with the Platinum Taq Hot Start Green DNA Polymerase and Platinum Green Hot Start PCR Master Mix (2X) compatible with downstream applications?

Yes the green dye is compatible with downstream applications such as fluorescent automatic DNA sequencing, ligation, and restriction digestion.

Do you offer Platinum Taq DNA Polymerase formats that already contain loading dyes in the reaction buffer?

Yes, we offer several Platinum Taq DNA polymerases and their master mixes, supplemented with tracking dyes for direct loading of PCR products on gels:

- Platinum II Hot-Start Green PCR Master Mix (2X)
- Platinum SuperFi Green DNA Polymerase and Platinum SuperFi Green PCR Master Mix
- Platinum Taq Green Hot-Start DNA Polymerase and Platinum Green Hot-Start PCR Master Mix (2X)
- DreamTaq Green Hot-Start DNA Polymerase and DreamTaq Hot-Start Green PCR Master Mix
- Phusion Green Hot-Start II High-Fidelity DNA Polymerase and Phusion Green Hot-Start II High-Fidelity PCR Master Mix.