AccuPower® HotStart Pfu PCR PreMix

AccuPower^® HotStart Pfu PCR PreMix is applied with Bioneer's enzyme-mediated HotStart patent technology to enhance response specificity and PCR amplification, along with reduction of non-specific reactions such as mis-priming and primer-dimer that may occure during Pfu DNA polymerase reaction at low temperatures. In addition, the proof-reading (3 '→ 5' exonuclease) function of Pfu DNA polymerase reduces the error in DNA amplification reaction, making it suitable for cloning experiments for protein production.

Furthermore, stabilizers are added to keep the activity stable for one month when stored at room temperature (25°C) and for two years when stored in a freezer.

※ This product is shipped at room temperature.
※ The reaction mixture contains stabilizers that keep full activity of enzymes even at the ambient temperature.

As low as $112.00

Cat. No.

Product Overview

Features and Benefits

Fidelity
High fidelity (error rate= 1.9X10^-6) with minimized mutation during DNA amplification
Specificity
Selection of specific sequences by applying PyroHotStart technology to the proof-reading function of Pfu DNA polymerase, allowing the amplified products to be used in experiments that require accurate sequences such as cloning and mutagenesis after simple PCR purification processes
User-friendly
Simplified procedure by requiring only the addition of template DNA, primer sets, and D.W., with all necessary components and DNA polymerases included and mixed in each of the tubes packaged with sufficient amount for 1 PCR run, along with application of Hotstart technology for amplifying specific targets accurately to minimize tedious steps such as Gel extraction for cloning
Stability
Stable enzyme activities allowing long-term storage by including stabilizer and dried in the PCR reaction mixture
Reproducibility

Minimized deviation between lots through a mass production under ISO 9001 quality system, allowing reproducible results even for replicates tested repeatedly under same conditions and variables

Application

• Gene cloning with blunt ends

• Site-directed mutagenesis

• High fidelity amplification

• High specificity PCR

• cDNA amplification

Experimental data

Figure. Comparison of PCR amplification efficiency between AccuPower^® Hotstart Pfu PCR PreMix from Bioneer and other suppliers' PCR master mix
Target : human insulin receptor gene. The cycling conditions for AccuPower^® Hotstart Pfu PCR PreMix were 95°C for 5 min, 30 cycles of 95°C for 30 sec, 55°C for 30 sec and 72°C for 2min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.

Lane M ; 100 bp DNA Ladder(Cat. No. D-1030,Bioneer)
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

Technical/Specs

▶ Specifications

Enzyme	Pfu DNA polymerase
5' to 3' exonuclease activity	No
3' to 5' exonuclease activity	Yes
3'- A overhang	No
Fragment Size	Up to 5 kb

▶ Experimental data

Figure 1. AccuPower^®Hotstart Pfu PCR PreMix shows enhanced specificity compared to competitors.
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, 32 cycles of 95°C for 30 sec, 62°C for 40 sec, and 72°C for 1 min 30sec, and 72°C for 5 min for final extension.

Lane M; 100 bp DNA Ladder (Cat. No. D-1030,Bioneer)
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 set1082 bp)
Lane 6; P65/83 primer set (1296 bp)
Lane 7; P55/83 primer set (1561 bp)

Figure 2. Comparison of PCR amplification specificity between AccuPower^® Hotstart Pfu PCR PreMix from Bioneer and other suppliers' pfu DNA polymerase.
Three different primer-template systems were amplified under the same conditions. The cycling conditions were 95°C for 5min, 35 cycles of 95°C for 30 sec, 62°C for 30 sec, and 72°C for 1 min 30 sec, and 72°C for 5 min for final extension.This data shows that AccuPower^® Hotstart Pfu PCR PreMix has higher amplificition efficency and specificity than other suppliers' pfu DNA polymerase.

Lane M; 100 bp DNA Ladder (Cat. No. D-1030,Bioneer)
Lane 1; 100 ng DNA, ApoE primer set (268 bp)
Lane 2; 10 ng DNA, ApoE primer set (268 bp)
Lane 3; 100 ng DNA, PrP primer set (500 bp)
Lane 4; 10 ng DNA, PrP primer set (500 bp)
Lane 5; 100 ng DNA, PrP primer set (705 bp)
Lane 6; 10 ng DNA, PrP primer set (705 bp)

Figure 3.AccuPower^® Hotstart Pfu PCR PreMix has high amplification efficiency.
Bioneer reaction mixture was followed by 95°C for 5min, 35 cycles of 95°C for 20 sec, 65°C for 20 sec, and 68°C for 15 min , and 68°C for 5 min for final extension.

Lane M; 1 kb DNA Ladder (Cat. No. D-1040,Bioneer)
Lane 1; 2 kb fragment
Lane 2; 2.5 kb fragment
Lane 3; 3 kb fragment
Lane 4; 4.5 kb fragment
Lane 5; 5 kb fragmentdirector: kim,hyunseo tel:578

Literature/Support

◈ Quality Assurance

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

• ISO 9001 - certificate

• ISO 13485 - certificate

Ordering Info

Cat. No.	Product Description	Option	Price
K-2301	AccuPower^® HotStart Pfu PCR PreMix	0.2 ml thin-wall 8-tube strips with attached caps /96 tubes, 20 μl reaction	$K-2301
K-2302		0.2 ml thin-wall 8-tube strips with attached caps /96 tubes, 50 μl reaction	$K-2302
K-2303		0.2 ml thin-wall 8-tube strips with attached caps /480 tubes,20 μl reaction	$K-2303
K-2304		0.2 ml thin-wall 8-tube strips with attached caps /480 tubes, 50 μl reaction	$K-2304