Master Mix High Fidelity (2X) | Master Mixes Cellco

Master Mix High Fidelity (2X)

Ficha técnica

Código	Master Mix High Fidelity (2X)
Categoria	PCR Convencional
Marca	Cellco

Descrição Geral

Master Mix High Fidelity (2X) | Master Mixes - Cellco
Master mix com polimerase termoestável de alta fidelidade
Pronto para uso

For in vitro use only!

Envio: Shipped on blue ice

Condições de armazenamento: Store at -20 °C

Avoid freeze/thaw cycles. Taq High Fidelity Pol – Master mix (2X) is also stable for three months at 4°C, so for frequent use, an aliquot may be kept at 4°C.

Validade: 24 months

Kit contents:

2x Taq High Fidelity Pol Master Mix (violet cap)
Master mix of thermostable DNA polymerase for high accuracy, dATP, dCTP, dGTP, dTTP, KCl, MgCl₂ and stabilizers.

Descrição:

Taq High Fidelity Pol Master Mix contains Taq High Fidelity Polymerase in an optimized PCR buffer with Mg²⁺ and dNTPs. It contains all reagents required for PCR (except template and primer) in a premixed 2x concentrated ready-to-use solution and should be used at a 1X concentration with DNA template and primers in a total reaction volume of 25 or 50 µL. The Master Mix is recommended for use in routine PCR reactions from templates including pure DNA solutions, bacterial colonies, and cDNA. It is optimized for high specificity and guarantees minimal byproduct formation.

PCR Reaction Setup:

Use the quantities below to prepare a single 50 µl PCR reaction. Thaw, mix, and briefly centrifuge each component before use. Add the following components to a microcentrifuge tube:

1. Prepare PCR master mix

Note: Consider the volumes for all components listed next steps to determine the correct amount of water required to reach your final reaction volume.

Component	50 μL rxn	Final conc.
Water, grade PCR	to 50 μl	-
2 X Taq High Fidelity Master Mix	25 μl	1x

Mix and briefly centrifuge the components.

2. Add template DNA and primers

Component	50 μl rxn	final conc.
Forward primer (10 μM)	0,5 – 2,5 μl	0,1 – 0,5 μM
Reverse primer (10 μM)	0,5 – 2,5 μl	0,1 – 0,5 μM
Template DNA	x μl	10 pg – 1 μg*

*genomic DNA: 1 ng – 1 μg; plasmidial DNA: 1 pg – 1 ng

Cap each tube, mix, and briefly centrifuge the content.

3. Incubate reactions in a thermal cycler

Recommended cycling conditions:

Initial denaturation	95 °C	1 - 3 min	1x
Denaturation	95 °C	15 - 30 sec	30 cycles
Annealing**	45 - 70 °C	15 - 30 sec	30 cycles
Elongation***	72 °C	1 min/kb	30 cycles
Final extension (optional)	72 °C	1 - 2 min	1x
Hold	4 – 10 °C	∞	1x

**The annealing temperature depends on the melting temperature of the primers used

***The elongation time depends on the length of the fragments to be amplified. A time of 1 min/kb is recommended.

Note: for large fragments it is recommended, in the elongation step, to use 68 ºC and 1,5 - 2 min/kbp

For optimal specificity and amplification an individual optimization of the recommended parameters may be necessary for each new template DNA and/or primer pair.

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