UltraPure™ Buffer-Saturated Phenol
UltraPure™ Buffer-Saturated Phenol
Invitrogen™

UltraPure™ Buffer-Saturated Phenol

El fenol saturado con tampón UltraPure™ se utiliza en la purificación de ácidos nucleicos. El reactivo, que consiste en fenolMás información
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Número de catálogoCantidad
15513039100 mL
15513047400 ml
Número de catálogo 15513039
Precio (MXN)
-
Cantidad:
100 mL
El fenol saturado con tampón UltraPure™ se utiliza en la purificación de ácidos nucleicos. El reactivo, que consiste en fenol UltraPure™ saturado con tampón Tris-HCl, ya está equilibrado a pH >7,4. Cuando se extraen mezclas con fenol saturado con tampón UltraPure™, las proteínas se desnaturalizan y se recogen en la fase orgánica o en la interfase, mientras que los ácidos nucleicos permanecen en la fase acuosa. El fenol saturado con tampón UltraPure™ no contiene conservantes. Se envasa en un gas inerte en botellas de color ámbar revestidas de plástico y resistentes a roturas.

Pruebas de rendimiento y calidad: El aspecto de la solución se evalúa a temperatura ambiente. No se ha detectado ninguna actividad de ribonucleasa (ARNasa) ni desoxirribonucleasa (ADNasa).
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Nombre del producto químico o materialFenoles
Tipo de empaquetadoFrasco
Línea de productosUltrapuro
PurezaGrado de biología molecular
Cantidad100 mL
Condiciones de envíoTemperatura ambiente
FormularioLíquido
pHpH 7,4
Unit SizeEach
Contenido y almacenamiento
Almacenar en el refrigerador (2–8° C).

Preguntas frecuentes

Do you have any information on DNA and RNA purification using phenol chloroform and alcohol precipitation?

Phenol extraction of proteins:

Phenol extraction is frequently used to remove proteins from nucleic acid solutions. A common protocol is to add an equal volume of buffer-saturated phenol or phenol:chloroform:isoamyl alcohol (25:24:1, v/v/v) to an aqueous nucleic acid solution, vortex, and centrifuge at 14,000 x g for 1 min to separate the phases.

Studies at Thermo Fisher Scientific have shown that the concentration of NaCl in the aqueous solution should not exceed 0.5 M for good recovery of DNA. Residual phenol can be removed from the aqueous phase by extraction with an equal volume of chloroform or ether. After extraction, DNA is usually precipitated with ammonium acetate and ethanol as described in another protocol on this server. Ref. Karger, B. D. (1989) FOCUS 11, 14.

A good source of general information on the properties of phenol can be found in Wallace, Donald M. “Large and Small-Scale Phenol Extractions”. Methods in Enz. Volume 152 guide to Molecular Cloning Techniques. 1987. Academic Press, Inc. Berger and Kimmel, eds. Chap.4, pg 33-41.

(a) At pH 5 to 6 DNA is selectively retained in the organic phase and interphase, leaving RNA in the aqueous phase. Therefore a pH greater than 7 is needed if DNA is to be extracted.

(b) At pH values below 7.6, poly A+ RNA is lost to the organic phase if chloroform is not present.

(c) Optimal RNA yields in phenol extraction are obtained if the salt concentration is less than 0.15 M NaCl. Salt concentration in the sample is not a factor for larger DNA molecules.

To store RNA after extraction use DEPC-treated water.

What is the recommended protocol for phenol-extraction removal of proteins from nucleic acid containing solutions?

Below is a commonly used protocol:

(1) Add an equal volume of buffer-saturated phenol or phenol:chloroform:isoamyl alcohol (25:24:1, v/v/v) to an aqueous nucleic acid solution. Note: for RNA solutions, acid-phenol is recommended.

(2) Vortex, and centrifuge at 14,000 x g for 1 min to separate the phases.

(3) Residual phenol can be removed from the aqueous phase by extraction with an equal volume of chloroform or ether.

(4) After extraction, DNA/RNA is usually precipitated with ammonium acetate and ethanol.

What is the optimal pH of the phenol:chloroform mixture for isolation of DNA?

Partitioning of the nucleic acids in phenol is pH dependent. At pH 7.0 or higher, both DNA and RNA partition into the aqueous phase. At an acidic pH (below 7.0) DNA is denatured and will move into the organic phase, but the RNA remains in the aqueous phase. The mixture should be adjusted to at least pH 7.4 for work with DNA.

Recently I came across a DNA purification technique, which uses urea during phenol extraction. What is the purpose of using urea?

Using urea during phenol extraction denatures the protein associated with the DNA and the proteins that bind the genomic DNA to the cell wall.