Platinum™ Hot Start PCR Master Mix (2X), 50 reactions - FAQs

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.
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.

Note: Addition of Platinum GC Enhancer solution enhances the amplification of GC-rich and problematic sequences.

The final concentration is 1.5 mM MgCl2.

We recommend using the GC enhancer for targets with >65% GC sequences. We recommend that you have a final concentration of 20% of the GC enhancer in the reaction mix. There is enough GC enhancer included in the kit to add it to every PCR reaction.

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.