SYBR™ Green Cells-to-C<sub>T</sub>™ Control Kit

Invitrogen™

SYBR™ Green Cells-to-C_T™ Control Kit

Dieses Kit ist für die Verwendung mit Power SYBR™ Green (SKU Nr. 4402953, 4402954 und 4402955) und Fast SYBR™ GreenWeitere Informationen

Katalognummer	Menge
4402959	100 Reaktionen

Katalognummer 4402959

Preis (EUR)

406,00

Menge:

100 Reaktionen

Preis (EUR)

406,00

Dieses Kit ist für die Verwendung mit Power SYBR™ Green (SKU Nr. 4402953, 4402954 und 4402955) und Fast SYBR™ Green Cells-to-C_T™ Kits (SKU Nr. 4402956 und 4402957). Es sind ausreichend Reagenzien für 100 Reaktionen enthalten. Es enthält Xeno™ RNA-Kontrolle, ein synthetisches RNA-Transkript mit einer einzigartigen Sequenz, die keine Homologie zu aktuellen annotierten biologischen Sequenzen aufweist, und ein PCR-Primer-Set für das Xeno™ RNA-Kontrollziel. Es enthält auch ein PCR-Primer-Set für das hoch exprimierte Gen β-Aktin (ACTB) für die endogene Kontrolle. Zusammen bieten diese Reagenzien positive Kontrollen für reverse Transkription und SYBR™ Green-basierte Echtzeit-PCR und können das Vorhandensein von reverse Transkriptions- oder PCR-Inhibitoren anzeigen. Echtzeit-RT-PCR mit SYBR™ACTB-Primern kann auch eine endogene Kontrolle für die Probennormierung bieten und eine ausreichende Zelleingabe in Experimenten mit Proben bestätigen, die aus mehr als 100 Zellen pro Lyse bestehen.

Nur für Forschungszwecke. Nicht zur Verwendung bei diagnostischen Verfahren.

Specifications

Kontroll-TemplateXenoRNA™ Kontrolle (synthetisches RNA-Transkript)

FormatTube

GensymbolACTB

Anzahl Reaktionen100 Reaktionen

ProduktlinieCells-to-CT, SYBR

Menge100 Reaktionen

VersandbedingungTrockeneis

ZielXenoRNA™-Kontrolle, β-Aktin

Marker oder FarbstoffUnkonjugiert

Unit SizeEach

Inhalt und Lagerung

Komponenten:

100 µl Xeno™ RNA-Kontrolle, 10^5 Kopien/µl

250 µl 20x SYBR Xeno Primer

250 µl 20x SYBR ACTB Primer

Mengen:

Xeno RNA-Kontrolle: ca. 100 Lysereaktionen

SYBR Xeno und SYBR ACTB Primer: bis zu 250 PCRs (je 20 µl)

Lagerungsbedingungen: Bei −20 °C lagern. Nicht in frostfreien Tiefkühlgeräten lagern.

Häufig gestellte Fragen (FAQ)

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