2×Super Pfx Master Mix

Cat. No.: S665634

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Overview

Products content

S665634	Component	1 mL	5 mL	Storage
S665634A	2×Super Pfx Master Mix	1 mL	5×1 mL	-20℃. Avoid freeze/thaw cycle.
S665634B	ddH₂O	1 mL	5×1 mL	-20℃. Avoid freeze/thaw cycle.

Products Introduction

It is an optimized 2× premixed reagent consisting of Super Pfx DNA Polymerase, Mg2+, dNTPs, and reaction buffer, and requires only the addition of primers and DNA templates for the PCR reaction.Super Pfx DNA Polymerase has a 3´→5´ exonuclease activity, and is approximately 100 times more fidelity than Taq DNA Polymerase. Super Pfx DNA Polymerase has 3´→5´ exonuclease activity and is about 100 times more fidelity than Taq DNA Polymerase, making it ideal for cloning. With the introduction of elongation enhancement technology, this enzyme enables high-speed PCR with an elongation speed of 10 sec/kb and an amplification length of up to 20 kb, while maintaining high fidelity. In addition, this product combines a high success rate with the ability to amplify long fragments that are difficult to amplify, GC-rich regions, and templates that are too large to be amplified. The amplified PCR product does not have an "A" base at the 3´ end and can be directly cloned into a flat-end vector. This product is divided into the standard 2×Master Mix without Loading Dye and the 2×Master Mix with Loading Dye.

Product Characteristics

1. High-speed PCR possible

The extension time can be set to 10sec for amplification of target fragments of less than 1kb, and 20-30sec/kb for amplification of target fragments of 1-10kb, which significantly reduces the PCR reaction time compared to conventional reagents.

2. High fidelity

The fidelity of Taq DNA polymerase is about 100 times higher than that of Taq DNA polymerase, and long target fragments can be amplified quickly and with high fidelity, and the amplified products can be used for a variety of purposes.

3. simple and convenient

This reagent contains all PCR components except primers and templates, which facilitates the operation and improves the reproducibility of the results. In addition, 2×Super Pfx Master Mix (Dye) contains Loading Dye, which can be used for gel electrophoresis directly after the reaction.

caveat

Primers and templates with uracil are not suitable for this product.

Instructions for use

PCR Reactors

1. Before preparing the reaction solution, please melt and mix the reagents completely before use.

After adding the components, please mix them well and transfer them to the PCR instrument quickly.

・The recommended final primer concentration is 0.4-0.5 μM, but for amplification of long fragments of 10 kb or more, a final primer concentration of 0.3 μM can increase the amount of amplified product.

PCR reaction program

1. Before preparing the reaction solution, please melt and mix the reagents completely before use.

take note of

1）Pre-denaturation: for most purified templates, 98℃, 30sec is sufficient; for complex templates, the pre-denaturation time can be extended, not more than 3min.

2）Annealing: In general, the annealing temperature is 3-5°C lower than the primer Tm. When a non-specific reaction occurs, increase the annealing temperature appropriately. If necessary, a temperature gradient can be established to find the optimal temperature for primer-template binding. For high Tm primers, a two-step cycle can be used, combining annealing and extension into one step.

3）Extension: for complex genomic samples, extension times are typically 20-30sec/kb, but can be reduced to 10sec/kb for simple templates (plasmids, E. coli, etc.) or complex templates <1kb. cDNAs, or complex templates >6kb, can be increased to 40-50sec/kb, if desired.

Specifications

Storage

Store at -20°C, Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.