2X M5 Taq HiFi PCR mix使用说明书

2X M5 Taq HiFi PCR mix使用说明书

产品名称 单位 货号 2×M5 Taq HiFi PCR Mix 1 ml 2×M5 Taq HiFi PCR Mix 5×1 ml 2×M5 Taq HiFi PCR Mix 10×1 ml

【储存条件】

长期保存，请置于-20˚C，有效期24个月。经常使用，可置于4˚C保存至少六个月。

MF002-01 MF002-05 MF002-10

【产品简介】

本产品包含高纯度M5 Taq HiFi DNA聚合酶、dNTPs、MgCl2、反应缓冲液、PCR反应的增强剂、优化剂以及稳定剂，浓度为2×。M5 Taq HiFi DNA 聚合酶比普通Taq酶扩增效率高、错配率低，具有快速简便、灵敏度高、特异性强、稳定性好等优点。可最大限度地减少人为误差，可用于高特异性PCR反应及GC含量较高（>60%）具有二级结构等复杂模板的扩增和大规模基因检测。PCR 产物的3’端附有一个突出的\"A\"碱基，纯化后可直接用于TA 载体克隆(货号MF019或MF020)。 本产品有含蓝色染料，在PCR 反应完成后，不需添加上样缓冲液即可直接上样进行电泳；也可经过纯化处理，以用于酶切、连接、荧光测序等后续操作。蓝色染料可指示电泳进程，其迁移速度在1% TAE琼脂糖凝胶中与500 bp 双链DNA 片段相近。

【产品组份】

2×M5 Taq HiFi PCR Mix 1 ml ddH2O 1ml

【适用范围】

1.基因检测：本产品不同批次之间误差很小，特别适合大规模基因检测、半定量PCR实验和微量DNA的检测。

2.用于从复杂模板中如基因组等扩增PCR产物，如表达基因的克隆、基因的定点突变和细胞内基因点突变的分析（SNP）等。

【所需试剂】使用者仅需准备PCR反应的模板、引物和蒸馏水等。

【操作示例】

按下表配制PCR反应体系（冰上操作）:

Template DNA*

<1µg

2× M5 Taq HiFi PCR Mix 1 0 μl Primer 1 (10 μM) 0.5 µl Primer 2 (10 μM) 0.5 µl

20 µl ddH2O补足至

建议的PCR条件： 95°C 32-36 cycles of: 94°C 55–°C 72°C 72°C 4°C

3 min. 25 sec. 25 sec. 30-60sec. /1 kb DNA 5 min. forever

【备注】

<*模板量：10~1000 ng 基因组 DNA，1~30 ng 质粒，或 1~2 μl RT-PCR 反应后的 cDNA。以上举例为常规 PCR 反应系统，仅供参考。实际反应条件因模板、引物等的结构不同而各异，需根据模板、引物、目的片段的特点设定最佳反应条件，并根据比例放大或缩小反应体系>。

本产品仅供科研使用。在确认产品质量出现问题时，本公司承诺为客户免费更换等量的质量合格产品。

2X M5 Taq HiFi PCR mix

Introduction

2× M5 Taq HiFi PCR mix is a convenient premixed 2× concentrated solution for PCR which includes M5 Taq HiFi DNA Polymerase,

2+

PCR Buffer, Mg, dNTPs, PCR Stabilizer and PCR Intensifier. Utilizing M5 Taq HiFi DNA polymerase with high amplification efficiency and low mismatching rate, the master mix offers the whole reaction system with high sensibility, specificity and stability. It is easy to use and can maximal avoid the personal error and contamination. It is suitable for PCR amplification with high specificity or large scale of gene test of difficult templates with secondary structure. The PCR product has adenine residue at the 3´-end of both strands, and it could be used for TA cloning (Mei5 Bio TOPO-TA cloning kit MF019 or MF020)、enzyme digestion、ligation and sequencing after purification. An inert blue dye (Bromophenol Blue, with migration equivalent to 500bp DNA fragment) is included in the master mix, which allows for direct loading of the PCR product(s) onto an agarose gel.

Components:

Cat. No. Description Quantity MF002-01 2× M5 Taq HiFi PCR Mix 1 ml MF002-05 2× M5 Taq HiFi PCR Mix 5×1 ml MF002-10 2× M5 Taq HiFi PCR Mix 10×1 ml Note: 2×M5 Taq HiFi PCR Mix contains Taq HiFi DNA Polymerase, 300 μM dNTP each, 20 mM Tris-HCl (pH8.7), 100 mM KCl, 3 mM MgCl2.

Shipping and Storage

Keep at –20°C for long-term storage. 2× M5 Taq HiFi PCR mix are stable at 4°C for six months. For daily use, we recommend keeping an aliquot at 4°C.

Applications:

1) Gene test, half- QPCR and detection of trace DNA.

2) Gene cloning, SNP analysis and site-directed mutagenesis.

Protocol

PCR reaction system Template DNA

<1µg

2× M5 Taq HiFi PCR Mix 1 0 μl Primer 1 (10 μM) 0.5 µl Primer 2 (10 μM) 0.5 µl ddH2O

PCR reaction conditions Procedure Pre-denaturation Denaturation Annealing Extension Final extension Keeping Temperature 95°C 94°C 55–°C 72°C 72°C 4°C Time 3 min. 25 sec. 25 sec. 30-60sec. /1 kb DNA 5 min. forever 32-36cycles Up to 20 µl