AccuPower® PyroHotStart Taq PCR PreMix

AccuPower® PyroHotStart Taq PCR PreMix는 바이오니아의 enzyme-mediated HotStart 특허기술을 적용하여 반응 특이성과 PCR 증폭효율을 높인 제품으로, Taq DNA polymerase가 낮은 온도에서 반응하면서 발생할 수 있는 mis-priming, primer-dimer와 같은 비특이 반응을 줄여줍니다. 그리고 DNA polymerase, dNTPs, reaction buffer 등 PCR 수행에 필요한 구성성분이 혼합되어 1회 분량씩 진공 건조되어 있습니다.

₩69,900
카탈로그 번호
K-2611-CFG

개요

(a) Zero cycle에서 PCR 반응 억제
PCR mixture 제조 후, pre-denaturation 온도에 도달하기 전에 비특이적 반응물이 생성되는 것을 방지하기 위해 PPi를 이용합니다.
DNA polymerase가 활성을 갖기 위해서는 Mg2+ 이온이 필요한데, mixture에 첨가되어 있는 PPi는 Mg2+ 이온과 높은 친화력을 갖기 때문에
Mg2+ 이온을 붙잡아 둠으로써 PCR 반응을 억제합니다.


(b) 변성 (Denaturation)
Mixture에 첨가되어 있는 PPase는 denaturation 단계에서 반응 온도가 올라가면 활성화되어 PPi를 Pi로 분해하여 Mg2+ 이온을 해리시킵니다.
해리된 Mg2+ 이온은 DNA polymerase에 결합하여 PCR 반응을 진행할 수 있도록 enzyme을 활성화 합니다.


(c) 풀림(Annealing) &증폭 (Extension)
활성화된 DNA polymerase는 PCR cycle이 진행됨에 따라 target DNA를 증폭합니다.

특장점

  • 특이성

    PyroHotStart 기술을 적용하여 적용하여 비특이 증폭산물 생성을 최소화하고, PCR 반응 효율을 극대화 함으로써 미량의 template DNA를 효과적으로 증폭할 수 있습니다.

  • 안정성

    PCR 반응 혼합물에 안정화제를 첨가하여 건조시켜, 장기간 보관하더라도 활성을 안정적으로 유지합니다.

  • 편의성

    각각의 PCR tube에 DNA polymerase와 PCR 수행에 필요한 모든 구성 성분이 포함되어 있어 template DNA, primer set, D.W.만 넣어 바로 PCR 반응을 수행할 수 있습니다. 또한 전기 영동 시에 필요한 tracking dye 와 침강제가 포함되어 있어 sample loading buffer를 추가로 넣을 필요가 없으므로 간편하게 사용할 수 있습니다.

  • 재현성

    ISO 9001 품질 시스템 하에서 one-batch system으로 대량 생산되어 각 batch에 대한 철저한 QC를 거친 후 균일한 품질의 제품으로 공급되기 때문에 사용자가 대량의 시료를 반복적으로 처리할 때 각 tube마다 발생하는 편차 문제를 해결하여 재현성있는 실험 결과를 제공합니다.

응용 및 적용

• High specificity PCR

• High sensitivity PCR

• gDNA template PCR

• Low-copy target PCR

• Multiple primer pairs PCR

• cDNA template PCR

• TA cloning

실험 자료

Figure. Comparison of PCR amplification specificity between AccuPower® PyroHotStart Taq PCR PreMix from Bioneer and other supplier's HotStart 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 amplificition efficency and specificity than other supplier's HotStart PCR master mix.

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

제품 규격/사양

Enzyme

Taq DNA polymerase

5' to 3' exonuclease activity

Yes

3' to 5' exonuclease activity

No

3'- A overhang

Yes

PCR product size

Up to 5 kb (human)


실험 자료

Figure 1. Comparison of PCR amplification specificity between AccuPower® PyroHotStart Taq PCR PreMix from Bioneer and other supplier's HotStart PCR master mix.
The ApoE gene was amplifed from 100 ng of human genomic DNA (The PCR product size is 268 bp). This data shows that AccuPower® PyroHotstart Taq PCR PreMix has higher amplificition efficency and specificity than other supplier's HotStart PCR master mix.

Lane M: 100 bp DNA Ladder (Cat. No.D-1030, Bioneer) 
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 2. 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; 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 set (1082 bp)
Lane 6; P65/83 primer set (1296 bp)
Lane 7; P55/83 primer set (1561 bp)




Figure 3. Comparison of PCR amplification between AccuPower® PyroHotStart Taq PCR PreMix from Bioneer and other supplier's HotStart 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 supplier's HotStart PCR master mix.

Lane M; 100 bp DNA Ladder (Cat. No.D-1030, Bioneer)
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 4. Comparison of cDNA template amplification between AccuPower® PyroHotStart Taq PCR PreMix and other supplier's HotStart 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 (Cat. No. K-2201, Bioneer) was used as a template for PCR amplification. This data shows that AccuPower® PyroHotstart Taq PCR PreMix has higher amplification efficiency than other supplier's HotStart PCR master mix.

Lane M; 100 bp DNA Ladder (Cat. No.D-1030, Bioneer)
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_AccuPower® PyroHotStart Taq PCR PreMix


MSDS

MSDS_AccuPower® PyroHotstart Taq PCR Premix


Brochure

AccuPower® PyroHotstart Taq PCR Premix

AccuPower® PyroHotStart Taq &Pfu PCR PreMix_Flyer

All about PCR


Quality Assurance

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

ISO 9001 - certificate

ISO 13485 - certificate

카탈로그 번호 제품명 가격
K-2611 AccuPower® PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached caps / 96 tubes, 20μl reaction/tube ₩69,900
K-2612 AccuPower® PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached caps / 480 tubes, 20μl reaction/tube ₩262,000
K-2613 AccuPower® PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached caps / 96 tubes, 50μl reaction/tube ₩105,000
K-2614 AccuPower® PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached caps / 480 tubes, 50μl reaction/tube ₩393,000
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