Cells-to-CT Bulk RT Reagents
Cells-to-C<sub>T</sub> Bulk RT Reagents
Invitrogen™

Cells-to-CT Bulk RT Reagents

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Los reactivos de RT a granel Cells-to-CT son versiones en gran cantidad disponibles por separado de los reactivos utilizados paraMás información
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Número de catálogoCantidad
4391852C2500 reactions
Número de catálogo 4391852C
Precio (MXN)
-
Cantidad:
2500 reactions
Los reactivos de RT a granel Cells-to-CT son versiones en gran cantidad disponibles por separado de los reactivos utilizados para la transcripción inversa en el kit de expresión génica Cells-to-CT™ TaqMan™ (mezcla de enzimas RT 20X y tampón RT 20X). Estos reactivos son suficientes para 2500 reacciones de RT. Cuando se combina con los reactivos de lisis a granel Cells-to- CT, estos reactivos pueden incorporarse fácilmente en aplicaciones automatizadas de alto rendimiento.
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Características ecológicasMenos recursos utilizados y menos residuos, menos peligroso
N.º de reacciones2500 reacciones
Línea de productosCells-to-CT
Tipo de productoReactivos de síntesis de ADNc
Cantidad2500 reactions
Condiciones de envíoHielo seco
Concentración20X
FormularioLíquido
Unit SizeEach
Contenido y almacenamiento
Mezcla de enzimas 20X RT, almacenar a –20 °C
Tampón 2X RT, almacenar a 4 °C

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.

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