Taq Pol (+)MgCl2 | Taq DNA Polimerase Cellco

Taq Pol (+)MgCl2

Ficha técnica

Código	Taq Pol (+)MgCl2
Categoria	PCR Convencional
Marca	Cellco

Descrição Geral

Taq Pol (+)MgCl₂ | Taq DNA Polimerase - Cellco

For in vitro use only!

Definição de unidade: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dNTPs into an acidinsoluble form in 30 minutes at 70 °C.

Envio: Shipped on blue ice

Condições de armazenamento: Store at -20 °C

Avoid freeze/thaw cycles

Validade: 12 months

Forma: Liquid

Concentração: 5 units/μL

Descrição:

Taq Pol is recommended for use in routine PCR reactions. It is optimized for high specificity and guarantees minimal byproduct formation. The buffer system is recommended for plate based PCR and automated pipetting. The enzyme replicates DNA at 72 °C. It catalyzes the polymerization of nucleotides into duplex DNA in 5' → 3' direction in the presence of magnesium. It also possesses a 5' → 3' polymerization-dependent exonuclease replacement activity but lacks a 3' → 5' exonuclease activity.

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 assay	Final conc.
PCR grade water	to 50 μl	-
10x Reaction Buffer Complete	5 μl	1x
dNTP Mix 10 mM	1 μl	200 μM
Taq DNA Polymerase (5 U/μl)	0,25-0,5 μl	1,25-2,5 U/assay
template DNA	x μl	<500 ng

Mix and briefly centrifuge the components.

2. Add template DNA and primers

Component	50 μl assay	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. Optimization of MgCl₂ concentration:

The 10x reaction buffer contain 20 mM MgCl₂, a recommended concentration for most applications. For an individual optimization add MgCl₂ stock solution as shown in the table below.

MgCl₂ Final Concentration	2,5 mM	3 mM	4 mM
MgCl₂ Stock volume to 50 μl	1 μl	2 μl	4 μl

4. 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 - 68 °C	15 - 30 sec	30 cycles
Elongation***	72 °C	1 min/kb	30 cycles
Final extension (optional)	72 °C	1 – 2 min/kb	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.

Produtos visitados

Nenhum produto visitado

Informações Detalhadas

Nosso tempo de faturamento é de até um dia útil durante nosso horário de funcionamento (Seg-Qui: 08:00 às 17:30 | Sex: 08:00 às 16:00). Caso a compra seja feita na sexta-feira, o faturamento só ocorrerá segunda-feira.

Produtos com disponibilidade IMEDIATA estão em nosso estoque próprio: Favor considerar 01 dia útil p/ faturamento + Tempo de transporte (a ser visualizado na página do produto)

Produtos que NÃO tenham disponibilidade IMEDIATA: Favor considerar Prazo em dias úteis informado em destaque + 01 dia útil p/ faturamento + Tempo de transporte (a ser visualizado na página do produto).