Century™-Plus RNA Markers

Los marcadores de ARN Ambion™ Century™-Plus son un conjunto de siete transcripciones in vitro de 100, 200, 300, 400, 500,Más información

Número de catálogo	Cantidad
AM7145	1 tubo

Número de catálogo AM7145

Precio (MXN)

Cantidad:

1 tubo

Pedido a granel o personalizado

Los marcadores de ARN Ambion™ Century™-Plus son un conjunto de siete transcripciones in vitro de 100, 200, 300, 400, 500, 750 y 1000 bases. No están etiquetados para su tinción con EtBr y se suministran en un tubo que contiene 50 µg a una concentración de 1 mg/ml. Se mezclan previamente para proporcionar una tinción equivalente en un gel de urea de poliacrilamida/8 M al 5 % y son útiles como estándares de tamaño de ARN en electroforesis en gel desnaturalizante. Los marcadores de 1–2 µg (2–4 µl) generarán siete bandas distintas en un gel de acrilamida TBE-Urea con tinción EtBr. Los marcadores también se prueban para estar libres de nucleasas y estables durante la noche a 37 °C.

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

Especificaciones

Concentración1 mg/ml

Compatibilidad del gelGeles de agarosa desnaturalizantes, geles de poliacrilamida desnaturalizantes

Masa50 µg

Línea de productosAmbion, siglo de ARN

Tipo de productoMarcador de ARN

Cantidad1 tubo

Listo para cargarNo

Condiciones de envíoHielo seco

Intervalo de tamañosDe 0,1 a 1 kb

Unit SizeEach

Contenido y almacenamiento

Almacenar por debajo de – 70 °C.

Preguntas frecuentes

Why are my RNA bands not sharp?

(1) RNA was not completely denatured. Electrophorese RNA under denaturing conditions. Use urea, formamide or formaldehyde gels, or glyoxal-treated RNA.

(2) For glyoxal-treated RNA, a buffer gradient formed during electrophoresis. Recirculate buffer during electrophoresis to prevent gradient formation.

What is the cause of extra bands when using an RNA ladder?

Extra bands appear in RNA Ladders for a few reasons:

(1) RNA was not completely denatured. Electrophorese RNA under denaturing conditions. Use urea, formamide or formaldehyde gels, or glyoxal-treated RNA.

(2) Extra bands may be a result of using formaldehyde that is not fresh; the pH becomes acidic in older formaldehyde. In our hands, when fresh formaldehyde with neutral pH was used, the extra bands disappeared.

(3) Alternatively, if the extra bands appear after hybridization, it could be that the gel purified probe contains contaminating vector DNA (pUC or pBR) that hybridizes to RNA Ladder template DNA.

Why are some of the RNA marker bands not visible?

Missing RNA bands may be due to:

(1) A small amount of RNA diffused out of gel during extended destaining. Minimize destaining time. Destaining for 2 hours is sufficient for most applications.

(2) RNA bands of similar molecular size were not resolved. Use the correct gel type and denaturing conditions.

Why are the RNA bands disappearing when looking and photographing a gel on a UV box?

RNA was exposed to UV light for extended periods of time. Minimize exposure to UV light. Stain and destain gels in the dark and photograph the gel immediately.

Why are the RNA marker bands so faint?

Many factors could affect the intensity of the bands as summarized below.

(1) Insufficient RNA was loaded on the gel. Increase the amount of RNA loaded.

(2) RNA was degraded. Avoid nuclease contamination of the RNA standards. Store RNA at -70 degrees C in formamide. Deionize formamide and glyoxal, store aliquots at -20 degrees C.

(3) RNA was electrophoresed off the gel. Electrophorese the gel for less time, at a lower voltage, or in a higher percentage gel.

(4) For ethidium bromide-stained RNA, improper UV light source was used. Use short-wavelength (254 nm) UV light. For ethidium bromide-stained RNA, improper staining and destaining conditions were used. Stain in the dark with 5 mg/mL ethidium bromide. For thin (3.2 mm) formaldehyde gels, stain 5 min and destain 1 h. For thicker gels, stain 30 min and destain 2 hr. To reduce background staining, use 0.66 M formaldehyde instead of 2.2 M formaldehyde in gels. For glyoxal RNA gels, stain and destain in 0.5 M ammonium acetate to help reduce background staining.

(5) For radiolabeled RNA, an improper labeling method was used.

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