AccuPower® HotStart PCR PreMix (with UDG)

AccuPower® HotStart PCR PreMix(with UDG)는 바이오니아의 enzyme-mediated HotStart 특허기술을 적용하여 반응 특이성과 PCR 증폭효율을 높였으며, uracil DNA glycosylase system을 적용하여 carry-over contamination 및 cross-over contamination 문제를 해결한 제품입니다. 그리고 DNA polymerase, dNTPs, reaction buffer 등 PCR 수행에 필요한 구성성분이 혼합되어 1 회 분량씩 진공 건조되어 있으며, 안정화 물질이 첨가되어 있어 상온 (25°C) 보관할 경우에는 한 달 동안, 냉동 보관 할 경우에는 2년 동안 활성이 안정적으로 유지됩니다.

₩102,000
카탈로그 번호
K-5050-1-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

  • 교차오염방지

    Uracil DNA glycosylase system 이 적용되어 Carryover contamination 에 의한 위양성을 최소화로 방지할 수 있습니다.

     accupower figure2

  • 민감도

    미량의 DNA 도 뛰어난 특이도와 높은 민감도로 PCR 반응이 가능합니다.

  • 편의성

    PyroHotstart 기술로 PCR 조건 선정 시간을 단축시킬 수 있으며 누구나 손쉽게 어려운 template DNA 도 타겟만을 정확히 얻을 수 있습니다.

  • 안정성

    PCR 반응 혼합액에 안정화제가 포함되어 있어 상온배송이 가능하고 solution type 제품보다 안정성이 뛰어납니다.

  • 재현성

    ISO 9001 품질 시스템 하에서 대량 생산되어 lot 간 편차가 최소화 되었으며, 반복적인 실험에도 재현성 있는 실험 결과를 제공합니다.

응용 및 적용

• gDNA template PCR

• Low-copy target PCR

• Multiple primer pairs PCR

• cDNA template PCR

• Molecular diagnosis

실험 자료

accupower

Figure. Comparison of specificity between AccuPower® HotStart PCR PreMix and HotStart PCR PreMix (with UDG).

Specificity test was operated using 7 pairs of primer targeting P53 gene. Reaction mixture was incubated at 37°C for 2 min followed by
95°C for 5 min, 30 cycles of 20 sec at 95°C, 40 sec at 55°C, 1 min at 72°C. AccuPower® HotStart PCR PreMix (with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP.

The amount of DNA (human) used to test is 10 ng and size of PCR amplicon size each lane is represented below.

Lane 1: 139 bp
Lane 2: 211 bp
Lane 3: 447 bp
Lane 4: 618 bp
Lane 5: 1082 bp
Lane 6: 1296 bp
Lane 7: 1561 bp
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)

제품 규격/사양

Enzyme

Top DNA polymerase

5' to 3' exonuclease activity

No

3' to 5' exonuclease activity

No

3'- A overhang

Yes

Fragment size

Up to 12 kb


실험 자료

accupower

Figure 1. Comparison of sensitivity between AccuPower® HotStart PCR PreMix and AccuPower® HotStart PCR PreMix(with UDG).

Sensitivity test was operated using serial diluted human genomic DNA. Reaction mixture was incubated at 37°C for 2 min followed by,
95°C for 5 min, 30 cycles of 20 sec at 95°C, 20 sec at 55°C, 30 sec at 72°C.
AccuPower® HotStart PCR PreMix(with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP.

The amount of DNA used to test is represented below.

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
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)




accupower

Figure 2. Comparison of amplification quality using PCR products (not including uracil base) between
AccuPower® HotStart PCR PreMix and AccuPower® HotStart PCR PreMix (with UDG).

Amplification quality test was operated using serial diluted PCR products not including uracil base. AccuPower® HotStart PCR PreMix was also tested for negative control. Reaction mixture was incubatedat 37°C for 2 min followed by 95°C for 5 min, 30 cycles of 20 sec at 95°C, 20 sec at 55°C, 30 sec at 72°C.
AccuPower® HotStart PCR PreMix (with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP.

The copy number of PCR products used to test are represented below.

Lane 1; 1011 copy 
Lane 2; 1010 copy
Lane 3; 10copy
Lane 4; 10copy 
Lane 5; 10copy
Lane 6; 10copy
Lane 7; 10copy
Lane 8; 10copy 
Lane N; No template control
M; 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)




accupower

Figure 3. Efficiency of uracil DNA glycosylase using PCR product (including uracil base).

Efficiency test of uracil DNA glycosylase was operated using serial diluted PCR products including uracil base.
AccuPower® HotStart PCR PreMix was also tested for negative control. Reaction mixture was incubated at 37°C for 2 min followed by 95°C for 5 min, 30 cycles of 20 sec at 95°C, 20 sec at 55°C, 30 sec at 72°C. AccuPower® HotStart PCR PreMix (with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP.

The copy number of PCR products used to test are represented below.

Lane 1; 1011 copy 
Lane 2; 1010 copy
Lane 3; 10copy
Lane 4; 10copy 
Lane 5; 10copy
Lane 6; 10copy
Lane 7; 10copy
Lane 8; 10copy 
Lane N; No template control
M; 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)

Manual

Manual_AccuPower® HotStart PCR PreMix(with UDG)


MSDS

MSDS_AccuPower® HotStart PCR PreMix(with UDG)


Brochure

AccuPower® HotStart 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-5050-1 AccuPower® HotStart PCR PreMix (with UDG), 0.2 ml thin-wall 8-strip tubes with attached cap / 96 tubes ₩102,000
K-5051-1 AccuPower® HotStart PCR PreMix (with UDG), 0.2 ml thin-wall 8-strip tubes with attached cap / 480 tubes ₩382,000
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