DNA Polymerase I

Name: DNA Polymerase I
SKU: 18010025
Price: 946.08 USD

ADN Polimerasa I es una ADN polimerasa con 5´→3´ y 3´→5´ actividades de exodesoxirribonucleasa. La ADN polimerasa I también incorporaMás información

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Número de catálogo	Cantidad
18010025	1000 U
18010017	250 U

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Número de catálogo 18010025

Precio (USD)

946,08

Each

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Cantidad:

1000 U

Precio (USD)

946,08

Each

Añadir al carro de la compra

ADN Polimerasa I es una ADN polimerasa con 5´→3´ y 3´→5´ actividades de exodesoxirribonucleasa. La ADN polimerasa I también incorpora nucleótidos biotinilados.

• Aplicaciones: ADNasa dependiente de I con translación de mellas, síntesis de segunda cadena en la clonación de ADNc, relleno de salientes de 5´
• Fuentepurificada a partir de E. coli que expresa el gen ADN polimerasa I en un plásmido

Pruebas de rendimiento y calidad
Ensayo de endodesoxiribonucleasa; se determinó la eficiencia de la ADNasa dependiente de I con translación de mellas.

Definición de la unidad
Una unidad incorpora 10 nMol de desoxirribonucleótido total en material precipitado en ácido en 30 min a 37 °C usando molde/primer.

Condiciones de reacción de la unidad
50 mM de fosfato de potasio (pH 7.0), 6,7 mM de MgCl₂, 1 mM de 2-mercaptoetanol, 80 µg/ml de plantilla/primer, 32 µM de dTTP, 69 nM de [³H]dTTP y enzima en 100 µl durante 30 min a 37 °C.

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones

DescripciónADN polimerasa

Actividad exonucleasa3'-5', 5'-3'

Inicio en calienteNo

PolimerasaDNA Polymerase I

Cantidad1000 U

Condiciones de envíoHielo húmedo

Unit SizeEach

Contenido y almacenamiento

1000 unidades de ADN polimerasa en un tampón de almacenamiento de 50 mM de fosfato potásico (pH 7,0), 100 mM de KCl, 1 mM de DTT y 50 % (v/v) de glicerol. Se suministra en 5–10 unidades/µl.

Almacenar entre - 5 °C y - 30 °C.

Preguntas frecuentes

Is there anything to prevent AmpliTaq Gold DNA polymerase from extending from the 3’ end of a TaqMan probe in a 5’ nuclease assay?

Yes. There is a phosphate group on the 3' end of all TaqMan probes that prevents such extension.

How does AmpliTaq Gold DNA Polymerase differ from AmpliTaq DNA Polymerase?

AmpliTaq Gold DNA Polymerase is a modified form of AmpliTaq DNA Polymerase that contains a proprietary chemical (or so-called hot start molecule) bound to the enzyme's active site. In order to activate the AmpliTaq Gold DNA Polymerase fully, we recommend an initial activation step of 95 degrees C for 10 min when using GeneAmp 10X PCR Buffer I and/or GeneAmp 10X PCR Buffer II and Mg in one of our thermal cyclers. When using GeneAmp 10X PCR Gold Buffer, activation time can be reduced to 5 minutes. Once activation is complete, you can proceed with your standard PCR cycling program (denaturing, annealing, extension, etc).

Does AmpliTaq Gold DNA Polymerase contain exonuclease (proofreading) activity?

No, AmpliTaq Gold DNA polymerase does not contain proofreading activity, however fidelity in PCR amplifications utilizing this enzyme may be improved. High fidelity can be achieved by: 1. Decreasing the final concentration of each nucleotide to 40-50 uM. 2. Using the lowest MgCl2 concentration possible. 3. Using less enzyme. 4. Decreasing extension times. 5. Using the highest annealing temperature possible. 6. Using as few cycles as possible.

Does the fidelity of AmpliTaq DNA Polymerase change in the presence of base analogs?

The fidelity of this PCR enzyme is affected in two ways. First, AmpliTaq DNA Polymerase typically binds to and incorporates base analogs less efficiently than conventional dNTPs, which means that polymerase activity is lower in reactions that contain base analogs. Second, the analog may pair with more than one conventional complementary template base, so the analog may be incorporated at an increased level compared to conventional dNTPs. For the best fidelity, we recommend that base analogs are included at low concentrations in the reaction.

What is the expected half life of AmpliTaq DNA Polymerase at 95 degrees C?

The half-life of AmpliTaq DNA Polymerase at 95 degrees C is 40 min. During PCR, the sample is only incubated at the programmed temperature for approximately 20 seconds. Therefore, the cycling half-life of AmpliTaq Gold at 95 degrees C is approximately 100 cycles.

Example: AmpliTaq DNA Polymerase experiences about 20 seconds at 95 degrees C per PCR cycle. The t_1/2 is at least 33 minutes; (35-40 min). Therefore, 33 min/20 sec/cycle = 100 cycles. 100 PCR cycles reduces enzyme activity by 50%.

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