SYBR™ Green Cells-to-C_T™ Control Kit

Este kit está diseñado para usarse con los kits Power SYBR™ Green (n.º SKU 4402953, 4402954 y 4402955) y FastMás información

Número de catálogo	Cantidad
4402959	100 reacciones

Número de catálogo 4402959

Precio (MXN)

Cantidad:

100 reacciones

Este kit está diseñado para usarse con los kits Power SYBR™ Green (n.º SKU 4402953, 4402954 y 4402955) y Fast SYBR™ Green Cells-to-C_T™(n.º SKU 4402956 y 4402957). Se incluyen reactivos suficientes para 100 reacciones. Incluye control de ARN Xeno™, una transcripción de ARN sintético con una secuencia única que carece de homología a las secuencias biológicas anotadas actuales, y un conjunto de cebadores para PCR para el objetivo de control de ARN Xeno™. También incluye un juego de cebadores de PCR para el gen de control endógeno altamente expresado β-actina (ACTB). Juntos, estos reactivos proporcionan controles positivos para la transcripción inversa y la PCR en tiempo real basada en SYBR™ Green, y pueden indicar la presencia de cualquier transcripción inversa o inhibidores de PCR. La RT-PCR en tiempo real con cebadores SYBR™ ACTB también pueden proporcionar un control endógeno para la normalización de muestras, así como para confirmar una entrada celular suficiente en experimentos con muestras formadas por más de 100 células por lisis.

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

Especificaciones

Molde controlControl de XenoRNA™ (transcripción de ARN sintético)

FormatoTubo

Símbolo de genACTB

Características ecológicasMenos recursos utilizados y menos residuos, menos peligroso

N.º de reacciones100 reacciones

Línea de productosCells-to-CT, SYBR

Cantidad100 reacciones

Condiciones de envíoHielo seco

ObjetivoControl XenoRNA™, β-actina

Etiqueta o tinteno conjugado

Unit SizeEach

Contenido y almacenamiento

Componentes:

100 µl de control de ARN Xeno™, 10^5 copias/µl

250 µl de cebadores SYBR 20X Xeno

250 µl de cebadores SYBR ACTB 20X Xeno

Cantidades:

control de ARN Xeno: aproximadamente 100 reacciones de lisis

Cebadores SYBR Xeno y ACTB: hasta 250 PCR (20 µl) cada uno

Condiciones de almacenamiento: Almacenar a −20 °C. No almacenar en un congelador sin escarcha.

Preguntas frecuentes

I'm seeing PCR products in the minus-RT control after performing my Cells-to-CT experiment. What does this mean?

If PCR products are seen in the minus-RT control reaction, but not in the no-template control, it indicates that genomic DNA remains in the sample and that genomic DNA was amplified in real-time PCR. Please follow the suggestions below:

- Ensure the DNase I is mixed thoroughly into the Lysis Solution.
- Use fewer cells per lysis reaction.
- Lyse cells using Lysis Solution that is at room temperature, and make sure that the lysis reaction occurs at room temperature.
You can also try increasing the incubation time of the lysis reaction to 8 minutes and/or using Lysis Solution that has been warmed up to 25 degrees C for cell lysis.

I'm getting PCR products in the no-template PCR control when performing a Cells-to-CT experiment. What could cause this?

PCR products in the no-template PCR control indicate that the sample is contaminated with DNA. More stringent steps need to be taken to control contamination.

I'm getting no PCR product or unexpected PCR products after performing a Cells-to-CT experiment. What could be the cause of this?

Please review the following possibilities and suggestions:

- A problem with adding or mixing the Stop Solution: ensure that the Stop Solution was added directly to the lysate, as components of the Lysis Solution may inhibit RT-PCR if not fully inactivated.
- The RNA was degraded: keep cells in PBS on ice before starting the cell lysis procedure.
- RNase in the sample was not completely inactivated: Too many cells could have been used or too much PBS left on the cells, diluting the lysis solution.
- The lysates sat too long before going to room temperature: Do not allow lysates to sit longer than 20 minutes at room temperature once the Stop Solution has been added.
- The sample does not contain the target RNA: Verify that the procedure is working by using the XenoRNA Control in the sample. Also check that your PCR primers can amplify your target under the PCR conditions you are using.

I ran out of stop solution for my Cells-to-CT experiment. Can I purchase it separately?

Yes, it is available in 1 mL aliquots (Cat. No. 4402960).

I have genomic DNA contamination in my Cells-to-CT reaction. How do I get rid of it?

1. Ensure that all medium is removed from the wells.
2. Wash with an equal volume of room temperature 1X PBS after the medium is removed.
3. Ensure that the reaction happens at room temperature (the lysis reaction may not reach room temperature if the plate is on ice, if the plate was quickly moved to the bench, or if a cold lysis solution was added).
4. Warm lysis solution to room temperature before adding to cells.
5. Allow the lysis reaction to proceed for 8 minutes at 25 degrees C.

View all SYBR™ Green Cells-to-C_T™ Control Kit FAQs

SYBR™ Green Cells-to-CT™ Control Kit