Kit de purificación Dynabeads™ mRNA DIRECT™
Kit de purificación Dynabeads™ mRNA DIRECT™
Invitrogen™

Kit de purificación Dynabeads™ mRNA DIRECT™

El kit de ARNm DIRECT™ de Dynabeads™ está diseñado para el aislamiento rápido y sencillo de ARN mensajero (ARNm) poliadeniladoMás información
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Número de catálogoCantidad
610115 mL
6101210 mL
Número de catálogo 61011
Precio (CLP)
1.161.490
Each
Añadir al carro de la compra
Cantidad:
5 mL
Precio (CLP)
1.161.490
Each
Añadir al carro de la compra
El kit de ARNm DIRECT™ de Dynabeads™ está diseñado para el aislamiento rápido y sencillo de ARN mensajero (ARNm) poliadenilado puro e intacto (poliA) directamente de lisado crudo de tejidos y células animales y vegetales.El ARNm aislado es adecuado para su uso en todas las aplicaciones posteriores.Ventajas del kit de ARNm DIRECT™ de Dynabeads™:

Rápido: un procedimiento de 15 minutos que produce ARNm puro intacto
Aislamiento de ARNm de alta pureza: la mejor opción previa a la síntesis de ADN complementaria (ADNc)
Sensibilidad del aislamiento de ARNm: permite la síntesis de ADNc y la construcción de bibliotecas de ADNc a partir de muestras de partida ultrapequeñas (permite la construcción de bibliotecas de ADNc a partir de una única célula)

Descripción general del sistema
El protocolo de aislamiento se basa en pares de base entre los residuos de poliA en el extremo 3' de la mayoría del ARNm y los residuos de oligo (dT)25 unidos covalentemente a la superficie del Dynabeads™.Otras especies de ARN que carecen de una cola poliA no se hibridan con los gránulos y se eliminan fácilmente.El ARN ribosómico, el ADN, las proteínas y las pequeñas moléculas de ARN (como ARN de transferencia, micro ARN y pequeños ARN nucleolares) no se unen a los gránulos y se descartan.Los agentes inhibidores de ARNasa en el tampón de lisis/unión junto con estrictas condiciones de lavado e hibridación garantizan el aislamiento de ARN puro intacto a partir de muestras crudas ricas en ARNasa, sin utilizar agentes caotrópicos fuertes.Los gránulos (beads) de purificación de ARNm se dirigen y capturan la transcriptoma de ARNm a partir de una amplísima variedad de muestras iniciales crudas (véase el protocolo).1 mg de gránulos Dynabeads™ oligo (dT)25 (200 μl) enlaza hasta 2 μg de ARNm.Una célula de mamífero típica contiene alrededor de entre 10 y 30 pg de ARN total, de los cuales entre un 1 % y un 5 % es ARNm.

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

Especificaciones
Tipo de producto finalARNm
Para utilizar con (aplicación)PCR cuantitativa en tiempo real (qPCR), PCR de transcriptasa inversa (RT-PCR), construcción de bibliotecas de ADNc, ensayos de protección contra nucleasas, northern blot, clonación
Compatibilidad de alto rendimientoCompatible con alto rendimiento, protocolos manuales y protocolos automatizados
Línea de productosDYNAL, Dynabeads, mRNA DIRECT
Tiempo de purificación15 min
Cantidad5 mL
Condiciones de envíoTemperatura ambiente
Isolation TechnologyGránulo magnético
Tipo de muestraSangre, células, FFPE y muestras fijas, muestras líquidas (p. ej., suero), muestras vegetales, ARN, tejido, muestras víricas, levadura, células, FFPE y muestras fijas, Muestras líquidas (p. ej., suero), Plant Samples, muestras vegetales, ARN, tejido, muestras víricas
TargetARNm
Unit SizeEach
Contenido y almacenamiento
Contenido:

5 ml de Dynabeads con oligo (dT)25

30 ml de tampón de unión/lisis

60 ml de tampón de lavado A

30 ml de tampón de lavado B

15 ml de Tris-HCl de 10 mM (tampón de elución)

Contiene reactivos suficientes para 20 aislamientos estándar. Almacenar a entre 2 y 8 °C.

Preguntas frecuentes

I am getting DNA contamination after mRNA isolation using Dynabeads magnetic beads. Why is this?

There are several reasons why DNA contamination may occur:

- Incomplete DNA shearing.
- Incomplete removal of sample lysate after the hybridization step.
- Insufficient washing and/or removal of wash buffers.
- The ratio of sample to beads was too high.

My Dynabeads magnetic beads are not pelleting well with the magnet. Do you have any suggestions for me?

Please review the following possibilities for why your Dynabeads magnetic beads are not pelleting:

- The solution is too viscous.
- The beads have formed aggregates because of protein-protein interaction.

Try these suggestions: - Increase separation time (leave tub on magnet for 2-5 minutes)
- Add DNase I to the lysate (~0.01 mg/mL)
- Increase the Tween 20 concentration to ~0.05% of the binding and/or washing buffer.
- Add up to 20 mM beta-merecaptoethanol to the binding and/or wash buffers.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

I have a long double-stranded DNA fragment I would like to isolate. What product do you recommend?

For biotin-labeled DNA that is less than 1 kb, we recommend you use Dynabeads M270 Streptavidin (Cat. No. 65305) and MyOne C1 magnetic beads (Cat. No. 65001). We recommend our Dynabeads KilobaseBINDER Kit (Cat. No. 60101), which is designed to immobilize long (>1 kb) double-stranded DNA molecules. The KilobaseBINDER reagent consists of M-280 Streptavidin-coupled Dynabeads magnetic beads along with a patented immobilization activator in the binding solution to bind to long, biotinylated DNA molecules for isolation. Please see the following link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/immobilisation-of-long-biotinylated-dna-fragments.html) for more information in regards to long biotinylated DNA fragment isolation.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

Can I use Dynabeads magnetic beads to isolate single-stranded DNA templates?

Yes, Dynabeads magnetic beads can be used to isolate single-stranded DNA. Streptavidin Dynabeads magnetic beads can be used to target biotinylated DNA fragments, followed by denaturation of the double-stranded DNA and removal of the non-biotinylated strand. The streptavidin-coupled Dynabeads magnetic beads will not inhibit any enzymatic activity. This enables further handling and manipulation of the bead-bound DNA directly on the solid phase. Please see the following link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/preparing-single-stranded-dna-templates.html) for more information in regards to single-stranded DNA capture.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

What is the magnetic susceptibility for Dynabeads magnetic beads?

Magnetic susceptibility is a measure of how quickly the beads will migrate to the magnet. This will depend on the iron content and the character of the iron oxide. The magnetic susceptibility given for the Dynabeads magnetic beads is the mass susceptibility, given either as cgs units/g or m^3/kg (the latter being an SI unit). For ferri- and ferromagnetic substances, the magnetic mass susceptibility is dependent upon the magnetic field strength (H), as the magnetization of such substances is not a linear function of H but approaches a saturation value with increasing field. For that reason, the magnetic mass susceptibility of the Dynabeads magnetic beads is determined by a standardized procedure under fixed conditions. The magnetic mass susceptibility given in our catalog is thus the SI unit. Conversion from Gaussian (cgs, emu) units into SI units for magnetic mass susceptibility is achieved by multiplying the Gaussian factor (emu/g or cgs/g) by 4 pi x 10^-3. The resulting unit is also called the rationalized magnetic mass susceptibility, which should be distinguished from the (SI) dimensionless magnetic susceptibility unit. In general, magnetic mass susceptibility is a measure of the force (Fz) influencing an object positioned in a nonhomogenous magnetic field. The magnetic mass susceptibility of the Dynabeads magnetic beads is measured by weighing a sample, and then subjecting the sample to a magnetic field of known strength. The weight (F1) is then measured, and compared to the weight of the sample when the magnetic field is turned off (F0). The susceptibility is then calculated as K x 10^-3 = [(F1-F0) x m x 0.335 x 10^6], where K is the mass susceptibility of the sample of mass m. The susceptibility is then converted to SI units.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.