Control negativo de ARNip Stealth™ RNAi, contenido medio de GC

El control negativo de ARN pequeño de interferencia de GC medio Stealth™ RNAi es uno de los tres controles negativosMás información

Número de catálogo	Cantidad
12935300	250 μl

Número de catálogo 12935300

Precio (MXN)

Cantidad:

250 μl

El control negativo de ARN pequeño de interferencia de GC medio Stealth™ RNAi es uno de los tres controles negativos que se utilizan con los experimentos con ARNip Stealth™. Estos controles constituyen un método excelente para medir el efecto de su dúplex de ARNip experimental Stealth™ RNAi con respecto al fondo. El kit de control negativo de ARNip Stealth™ RNAi contiene los tres controles (contenido bajo, medio y alto de GC: sin incluir las secuencias n.º 2 y n.º 3), cada uno de los cuales está también disponible por separado. Estos controles:

• Diseñado con tres niveles de contenido de GC (bajo, medio, alto) con tres secuencias disponibles por nivel para que los investigadores puedan comparar el contenido de GC con su ARNip experimental Stealth™ RNAi
• No es homólogo a ningún elemento del transcriptoma de vertebrados
• Probado para no inducir una respuesta de estrés

Acerca de los controles negativos Stealth™ RNAi
Los dúplex de control negativo Stealth™ RNAi son perfectos para usar en experimentos de interferencia de ARN (ARNi) como control de los efectos independientes de la secuencia que se producen tras la administración de Stealth™ RNAi en la línea de células de cualquier vertebrado. Cada dúplex de controles negativos Stealth™ RNAi está diseñado para minimizar la homología de secuencia con cualquier transcrito de vertebrados conocido. Los dúplex de controles negativos Stealth™ RNAi se diferencian en el contenido de GC y se suministran en un formato listo para su uso. Si así se desea, se puede incluir un tampón de dilución/recocido de ARN para la dilución de la solución madre Stealth™ RNAi.

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

Especificaciones

Para utilizar con (aplicación)Transfección, ARNi

Etiqueta o tinteno conjugado

Línea de productosARNip Stealth RNAi

Tipo de productoARNip

Cantidad250 μl

Tipo de controlControl negativo

RNAi TypeARNip

Unit SizeEach

Contenido y almacenamiento

Contiene un tubo de dúplex de control negativo Stealth™ RNAi de 250 µl (20 µM) y un tubo de tampón de recocido/dilución de ARN 1X de 1 ml. Almacenar a -20 °C.

Preguntas frecuentes

What are the benefits of using a vector to deliver RNAi?

Vector technologies allow you to:

Achieve transient or stable target knockdown
Perform RNAi in any cell type, even hard-to-transfect, primary, and non-dividing cells
Regulate gene inhibition with inducible siRNA expression
Select for a pure population of cells stably expressing an siRNA sequence
Control gene expression in vivo with tissue-specific promoters

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

Why are my cells dying after transfection?

We would suggest running a transfection reagent control only to determine if your cells are sensitive to the transfection reagent. Additionally, you can try using different cell densities and siRNA concentrations to diminish any toxic effects from the transfection itself.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

I transfected my siRNA and the mRNA levels are down, but the protein is not. Why is that?

In some cases, knockdown of a protein can be affected by other variables such as protein turnover rate, even though the RNA is knocked down. Additionally, a longer time course may be needed to see an effect on protein compared to mRNA.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

I am not getting my target knockdown. What could be the cause of this?

Please see the following possibilities and suggestions:

- How many siRNA did you test? Is there any knockdown? If there is no knockdown (<10%) in any of the siRNA, then the assay is likely the problem. Try using a different qRT-PCR assay to assess knockdown.
- What was the positioning of the qRT-PCR assay target site relative to the cut site for the siRNA? If greater than 3,000 bases away, the problem could be alternative splice transcripts.
- What are the Cts for the experiment? They should be below 35 in a 40-cycle qRT-PCR experiment.
- Did you confirm the siRNA got into the cell? We recommend using a validated positive control siRNA to check the transfection efficiency.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

I am not getting any knockdown with my siRNA. What do you suggest I try?

Please see the following possibilities and suggestions:

- Were the mRNA levels checked? The most reliable method is real-time PCR. In some cases, knockdown of a protein can be affected by other variables, such as protein turnover rate, even though the RNA is knocked down.
- How is the RNA being isolated? Has the quality of the isolated RNA been checked? Ensure that the RNA has not been degraded.
- Was a positive control used? This can help to determine whether the reagents are working and whether the siRNA was delivered correctly to the cell. Run your experiment in parallel with the positive control siRNA.
- Was a transfection control used? What is the percentage of transfected cells?
- Was a time course used? Generally, gene silencing can be assessed as early as 24 hours posttransfection. However, the duration and level of knockdown are dependent on cell type and concentration of siRNA.
- Was optimization of transfection conditions performed? You can try using different cell densities and siRNA concentrations.
- Which concentration of siRNA did you use? We recommend testing multiple concentrations between 5 nM and 100 nM.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

View all Control negativo de ARNip Stealth™ RNAi, contenido medio de GC FAQs