柏业贸易

特点及优势

多重PCR
一个试管中仅需一个反应即可生成20种多重扩增产物
特异性
通过使用PyroHotStart技术（包括焦磷酸酶）将非特异性扩增减至最少，从而通过消除每个循环中产生的PCR抑制剂焦磷酸盐来最大程度地提高效率
便利性
PyroHotstart技术可以缩短多重PCR条件设定时间，任何人都可以通过仅添加模板DNA和几种类型的引物轻松地将多个模板从1扩增到20个
稳定性
添加了稳定剂可以在常温下运输，比溶液型产品更稳定
重现性
ISO 9001质量体系下的批量生产可最大程度地减少批次之间的差异，即使在重复实验中也可得到可重复的实验结果

概述

(a)在Zero cycle抑制PCR反应
As PPi has high affinity with Mg2+, 制备PCR混合液后，使用PPi防止在达到预变性温度之前形成非特异性反应物。为了使DNA聚合酶具有活性，需要镁离子，因为添加到混合液中的PPi与镁离子具有高亲和力，可以通过保留镁离子来抑制PCR反应。

变性(Denaturation)
在变性步骤中，当反应温度升高时，添加到混合液中的PPase会被激活，从而将PPi分解为Pi从而解离镁离子。解离的镁离子与DNA聚合酶结合并激活该酶催化PCR反应。

应用

实验数据

Figure. High specificity of AccuPower® Gold Multiplex PCR PreMix
Each lane from left to right represensts the progressive number of primer sets (1 – 20) included in single AccuPower® Gold Multiplex PCR PreMix reactions.
Rxn. Condition : 95°C for 10 min, followed by 30 cycles(a), 35 cycles(b) of 30 sec at 95°C, 30 sec at 57°C, 60 sec at 72°C
M; 25/100 bp Mixed DNA Ladder (Cat. No. D-1020, Bioneer

▶ Specifications

Enzyme	Top DNA polymerase
5' to 3' exonuclease	No
3' to 5' exonuclease	No
3' – A overhang	Yes
Fragment Size	Up to 1 kb

▶ Experimental data

Figure 1. Single PCR and Multiplex PCR using AccuPower^® Gold Multiplex PCR PreMix
Each lane from left to right indicates the single and multiplex PCR products using AccuPower^® Gold Multiplex PCR PreMix.
The last lane labeled multi represents all amplicons amplified in a single reaction.

a) 10 plex multiplex PCR
b) 20 plex multiplex PCR M; 25/100 bp Mixed DNA Ladder (cat. No. D-1020, Bioneer)

Figure 2. Comparison of amplification quality, using a Caliper Labchip^® 90 system, between AccuPower^® Gold Multiplex PCR PreMix and competitor Multiplex PCR kits.

a) Virtual Gel Image. Gel image illustrates data reproducibility of the Caliper LabChip^® 90 system.
b) Overlay of expression levels using Bioneer's AccuPower^® Gold Multiplex PCR PreMix and competitor multiplex PCR kits. The electropherogram displays the data between 10 PCR product yields using a 10-plex primer set to illustrate amplification efficiency.
c) The graph shows the total concentration of PCR products between AccuPower^® Gold Multiplex PCR PreMix and other supplier's master mix.

Figure 3. Comparison of amplification quality between AccuPower^® Gold Multiplex PCR PreMix and competitor Multiplex PCR kits.
6-plex (a), 10-plex (b), 20-plex (c) primers were added into AccuPower^® Gold Multiplex PCR PreMix and competitor master mix.
A series of human genomic DNA diluents were tested (Lane 1; 100 ng, Lane 2; 10 ng, Lane 3; 1 ng).
All data were obtained using Mygenie 96 machine (Applied Bioneer co.)

Supplier Q : Multiplex PCR master mix
Supplier S : Multiplex PCR master mix
Supplier I : Taq polymerase for Mutiplex PCR (0.5U), added 2 mM MgCl₂
M; 25/100 bp Mixed DNA Ladder (cat. No. D-1020, Bioneer)