Platinum™ SuperFi II DNA 聚合酶

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

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

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.

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