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Wednesday, 29 May 2013 16:18

AccuPower ProFi Taq PCR PreMix

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AccuPower® ProFi Taq PCR PreMix for high efficiency and amplification of long range PCR.

ProFiTaqPCRPreMix f01

AccuPower® ProFi Taq PCR PreMix is a convenient lyophilized PCR master mix containing ProFi Taq DNA polymerase, reaction buffer, dNTPs, tracking dye, and a patented stabilizer. ProFi Taq DNA polymerase in the premix is a unique recombinant Taq DNA polymerase that offers enhanced amplification efficiency and higher fidelity for PCR. AccuPower® ProFi Taq PCR PreMix is applicable to any template DNA, and especially effective in amplifying large genomic DNA fragments around 20 kb. AccuPower® ProFi Taq PCR PreMix provides accurate long-range amplification of standard and amplification of low-copy target, and is highly suitable for all PCR applications.

 

Features and Benefits

Long PCR: ProFi Taq is especially effective in amplifying large genomic DNA fragments around 20 kb and amplifying Lambda DNA up to 30kb.
Easy to use: All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized "PreMix" form.
Reproducibility: Gentaur's strict quality controlled production system ensures that your results will be reproducible experiment after experiment.
Convenient: Just add template and primers and start your reaction. dNTPs, buffer and enzyme are provided
Stability: Stable at room temperature for a month and for 2 years in a -20°C freezer

 

Specifications

5' to 3' exonuclease: Yes
3' to 5' exonuclease: Yes
3' – A Overhang: Yes
PCR product size: ~ 30kb


Application

- Primer extension
- long-range amplification from genomic DNA
- High amplification efficiency
- Excellent performance on difficult templates
- Amplification of low-copy targets
- High yield and high sensitivity PCR

Experimental data

ProFi Taq fig1

Figure 1. Comparison of PCR amplification efficiency between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5min, 30 cycles of 95°C for 20 sec, 55°C for 20 sec and 72°C for 30 sec. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Target : human insulin receptor gene.
Lane M : 100 bp DNA Ladder(Gentaur, Cat. No. D-1030)
Lane 1 : 10 ng of human genomic DNA
Lane 2 : 1 ng of human genomic DNA
Lane 3 : 100 pg of human genomic DNA
Lane 4 : 10 pg of human genomic DNA

ProFi Taq fig2
Figure 2. Comparison of PCR amplification efficiency between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

cDNA synthesized from 10-fold serial-diluted human total RNA from 10 ng to 10 pg using AccuPower® RocketScript™ Cycle RT PreMix. (Gentaur, Cat. No K-2201) wase used as a template for PCR amplification. The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5min, 33 cycles of 95°C for 20 sec, 55°C for 20 sec and 72°C for 30 sec. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Target : human GAPDH gene.
Lane M : 100 bp DNA Ladder(Gentaur, Cat. No. D-1030)
Lane 1 : 10 ng of human total cDNA
Lane 2 : 1 ng of human total cDNA
Lane 3 : 100 pg of human total cDNA
Lane 4 : 10 pg of human total cDNA

ProFi Taq fig3
Figure 3. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 30 cycles of 95°C for 20 sec, 65°C for 20 sec and 68°C for 4 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Lane M1 : Lambda/Hind III marker (Gentaur, Cat. No. D-1050)
Lane M2 : 1 kb DNA Ladder (Gentaur, Cat. No. D-1040)
Lane 1 : 2 kb fragment (human tumor protein p53 gene)
Lane 2 : 3 kb fragment (human tumor protein p53 gene)
Lane 3 : 4.5kb fragment (human DNA cross-link repair 1A gene)
Lane 4 : 8 kb fragment (human hemoglobin epsilon 1 gene)

ProFi Taq fig4
Figure 4. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 32 cycles of 95°C for 20 sec and 68°C for 15 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol. Human genomic DNA was used as a template for PCR amplification.
Lane M1 : Lambda/Hind III marker (Gentaur, Cat. No. D-1050)
Lane M2 : 1 kb DNA Ladder (Gentaur, Cat. No. D-1040)
Lane 1 : 11 kb fragment
Lane 2 : 13.5 kb fragment
Lane 3 : 17.6 kb fragment
Lane 4 : 21.4 kb fragment

ProFi Taq fig5
Figure 5. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 32 cycles of 95°C for 20 sec, 65°C for40 sec, and 68°C for 20 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol. Lambda DNA was used as a template for PCR amplification.
Lane M1 : Lambda/Hind III marker (Gentaur, Cat. No. D-1050)
Lane M2 : 1 kb DNA Ladder (Gentaur, Cat. No. D-1040)
Lane 1 : 15 kb fragment
Lane 2 : 20 kb fragment
Lane 3 : 25 kb fragment
Lane 4 : 30 kb fragment

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