DNA Polymerase I

Citas y referencias (2)

Invitrogen™

DNA Polymerase I

Name: DNA Polymerase I
SKU: 18010017

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
18010017	250 U
18010025	1000 U

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Número de catálogo 18010017

Precio (MXN)

Cantidad:

250 U

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

Cantidad250 U

Condiciones de envíoHielo húmedo

FormularioLíquido

Unit SizeEach

Contenido y almacenamiento

Almacenar en congelador (de -5 a -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%.

View all DNA Polymerase I, 250 U FAQs

Citations & References (2)

Citations & References

Abstract

Both DNA and histone fold sequences contribute to archaeal nucleosome stability.

Authors: Bailey Kathryn A; Marc Frederic; Sandman Kathleen; Reeve John N;

Journal:J Biol Chem

PubMed ID:11751933

'The roles and interdependence of DNA sequence and archaeal histone fold structure in determining archaeal nucleosome stability and positioning have been determined and quantitated. The presence of four tandem copies of TTTAAAGCCG in the polylinker region of pLITMUS28 resulted in a DNA molecule with increased affinity (DeltaDeltaG of approximately 700 ... More

Escherichia coli apurinic-apyrimidinic endonucleases enhance the turnover of the adenine glycosylase MutY with G:A substrates.

Authors: Pope Mary Ann; Porello Silvia L; David Sheila S;

Journal:J Biol Chem

PubMed ID:11960995

The DNA repair enzyme MutY plays an important role in the prevention of DNA mutations resulting from the presence of the oxidatively damaged lesion 7,8-dihydro-8-oxo-2'-deoxyguanosine (OG). MutY is a base excision repair (BER) glycosylase that removes misincorporated adenine residues from OG:A mispairs, as well as G:A and C:A mispairs. We ... More