Master mix Taq Pol (2X) | Master Mixes Cellco

Master mix Taq Pol (2X)

Ficha técnica

Código	Master mix Taq Pol (2X)
Categoria	PCR Convencional
Marca	Cellco

Descrição Geral

Master mix Taq Pol (2X) | Master Mixes - Cellco
Master mix de DNA polimerase termoestável
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 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: 12 months

Kit contents:

2x Taq Pol Master Mix (violet cap)
Composition: 0.05 U/ µL Taq DNA polymerase, reaction buffer, 0.3 mM MgCl₂, 0.4 mM of each dNTP (dATP, dCTP, dGTP, dTTP), and stabilizers.

Descrição:

Taq Pol Master Mix contains Taq 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 by-product formation.

PCR Reaction Setup:

The PCR procedure below shows appropriate volumes for a single 50-μL reaction. For multiple reactions, prepare a master mix of components common to all and then dispense appropriate volumes into each PCR reaction tube prior to adding template DNA and primers. 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 Pol 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 min	1x
Denaturation	95 °C	15 - 30 sec	30 cycles
Annealing**	45 - 68 °C	15 - 30 sec	30 cycles
Elongation***	72 °C	30 sec - 4 min	30 cycles
Final extension (optional)	72 °C	2 min	1x
Hold	4 – 8 °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.

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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