AccuPower® PyroHotstart Taq PCR Premix

Bioneer own patented DNA Amplification product with high specificit y.

AccuPower® PyroHotstart Taq PCR PreMix is a PCR master mix containing a thermostable DNA polymerase, thermostable pyrophosphatase, reaction buffer, dNTPs, tracking dye, and patented stabilizer in a ready to use hotstart PCR master mix. Bioneer uses a unique enzyme-mediated HotStart PCR that provides robust and reliable results. Bioneer's Taq DNA Polymerase is inhibited at lower temperatures (< 70°C) by pyrophosphate. However, Taq DNA Polymerase is rendered fully active at temperatures above 70°C via pyrophosphate hydrolysis with our thermostable pyrophosphatase. This prevents the formation of mis-primed products and primer-dimers during the reaction set up process resulting in improved PCR specificity. It is ideal for nucleic acid amplification reactions involving complex genomic or cDNA templates, very low copy targets, and multiplex reactions.
 

Features and Benefits

• Specificity

Pyrophosphate (PPi) has high affinity for Mg2+. By adding PPi to the reaction mixture, the Mg2+ ions necessary for normal PCR are bound, preventing DNA polymerase activity. This PPi-Mg2+ binding prevents non-specific before PCR (zero-cycle) product formation. Upon thermal cycling, the pyrophosphatase (PPase) that is also added to the mixture is activated (>70°C) and hydrolyzes the PPi to 2 phosphate groups and facilitates the release of Mg2+, which is then available for DNA polymerase to use and resume normal activity

    
 

• Stability

Stable at room temperature for a month and for 2 years in a -20°C freezer
 

• Ease-of-Use

All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized "PreMix" form. The user needs only to add template DNA, primers and water. Materials necessary for loading agarose gels for electrophoresis are also added in the reaction, negating the need to add loading dye after PCR is completed.
 

• Reproducibility

Each batch is produced under strict quality controls. Errors that commonly occur during mass production are eliminated during the individual packaging process. Bioneer’s current batch processing system allows for the production of more accurate and reproducible end-product yield

 

Specifications

5' to 3' exonuclease: Yes
3' to 5' exonuclease: No
3' – A Overhang: Yes
PCR product size: ~ 5 kb(Human)

 

Application

- High specificity PCR
- High sensitivity PCR
- Low-copy target PCR
- Multiple primer pairs PCR
- cDNA template PCR
- T-A cloning

 

Experimental data

Figure 1. Comparison of PCR amplification specificity between AccuPower® PyroHotStart Taq PCR PreMix from Bioneer and other suppliers' Hot start PCR master mix.

PCR reactions were performed according to each supplier's protocol. The PrP gene was amplified from human genomic DNA with two different primer sets, separately. This data shows that AccuPower® PyroHotStart Taq PCR PreMix has higher amplification efficiency and specificity than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder(Bioneer, Cat. No.D-1030)
Lane 1: 100 ng DNA, PrP primer set (500 bp)
Lane 2: 10 ng DNA, PrP primer set (500 bp)
Lane 3: 100 ng DNA, PrP primer set (705 bp)
Lane 4: 10 ng DNA, PrP primer set (705 bp)

Figure 2. Comparison of PCR amplification specificity between AccuPower® PyroHotStart Taq PCR PreMix from Bioneer and other suppliers' Hot start PCR master mix.

The ApoE gene was amplifed from 100 ng of human genomic DNA (The PCR product size is 268bp). This data shows that AccuPower® PyroHotStart Taq PCR PreMix has higher amplificition efficency and specificity than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder (Bioneer, Cat. No. D-1030)
Lane 1: AccuPower® PyroHotStart Taq PCR PreMix
Lane 2: Supplier I Hotstart Taq PCR preMix
Lane 3: Supplier S Hotstart Taq PCR masterMix
Lane 4: Supplier T Hotstart Taq PCR masterMix
Lane 5: Supplier Q Hotstart Taq PCR masterMix

Figure 3. AccuPower® PyroHotStart Taq PCR PreMix has high amplification efficiency and specificity.

Specificity test was performed using 7 different sets of primers targeting the P53 gene. 10 ng of human genomic DNA was used for each PCR reaction. The cycling conditions were 95°C for 5min, 30 cycles of 95°C for 20 sec, 55°C for 40 sec, and 72°C for 1 min, and 72°C for 5 min for final extension.
Lane M: 100bp DNA Ladder (Bioneer, Cat. No. D-1030)
Lane 1: P75/73 primer set (139 bp)
Lane 2: P55/53 primer set (211 bp)
Lane 3: P55/63 primer set (447 bp)
Lane 4: P75/83 primer set (618 bp)
Lane 5: P55/73 primer set (1082 bp)
Lane 6: P65/83 primer set (1296 bp)
Lane 7: P55/83 primer set (1561 bp)

Figure 4. Comparison of PCR amplification between AccuPower® PyroHotStart Taq PCR PreMix from Bioneer and other suppliers' Hot start PCR master mix.

Sensitivity test was performed by amplifying the IRGC gene from a serial dilution of human genomic DNA. This data shows that AccuPower® PyroHotStart Taq PCR PreMix has higher amplification efficiency than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder (Bioneer, Cat. No. D-1030)
Lane 1: 10 ng human genomic DNA
Lane 2: 1 ng human genomic DNA
Lane 3: 100 pg human genomic DNA
Lane 4: 10 pg human genomic DNA

Figure 5. Comparison of cDNA template amplification between AccuPower® PyroHotStart Taq PCR PreMix and other suppliers' Hot start 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 (Bioneer, Cat. No. K-2201) was used as a template for PCR amplification. This data shows that AccuPower® PyroHotStart Taq PCR PreMix has higher amplification efficiency than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder (Bioneer, Cat. No. D-1030)
Lane 1: 10 ng human total cDNA
Lane 2: 1 ng human total cDNA
Lane 3: 100 pg human total cDNA
Lane 4: 10 pg human total cDNA

 

Manual
• manual-PyroHotstart-Taq-PCR-PreMix

MSDS
• AccuPower® PyroHotstart Taq PCR PreMix

Brochure
• AccuPower® PreMix series - 2010 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

Cat.No.	Product Description	Price	Add to Cart
K-2611	AccuPower® PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-strip tubes with attached caps / 96 tubes, 20ul reaction/tube	Inquire
K-2612	AccuPower® PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-strip tubes with attached caps / 480 tubes, 20ul reaction/tube	Inquire
K-2613	AccuPower® PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-strip tubes with attached caps / 96 tubes, 50ul reaction/tube	Inquire
K-2614	AccuPower® PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall tubes with attached caps / 480 tubes, 50ul reaction/tube	Inquire