Platinum™ SuperFi II Green PCR 预混液

Typically, for a 50 µL PCR reaction, we recommend using 25µµL of the 2X master mix so that its final concentration in the reaction is 1X.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

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.

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

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

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.