SYBR™ Green Cells-to-CT™ Control Kit
SYBR&trade; Green Cells-to-C<sub>T</sub>&trade; Control Kit
Invitrogen™

SYBR™ Green Cells-to-CT™ Control Kit

Green features
This kit is designed for use with the Power SYBR™ Green (SKU#s 4402953, 4402954, and 4402955) and Fast SYBR™ GreenRead more
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Catalog NumberQuantity
4402959100 Reactions
Catalog number 4402959
Price (CNY)
4,507.00
Each
Add to cart
Quantity:
100 Reactions
Price (CNY)
4,507.00
Each
Add to cart
This kit is designed for use with the Power SYBR™ Green (SKU#s 4402953, 4402954, and 4402955) and Fast SYBR™ Green Cells-to-CT™ Kits (SKU#s #4402956 and 4402957). Sufficient reagents are provided for 100 reactions. It includes Xeno™ RNA Control, a synthetic RNA transcript with a unique sequence that lacks homology to current annotated biological sequences, and a PCR primer set for the Xeno™ RNA Control target. It also includes a PCR primer set for the highly expressed endogenous control gene β-actin (ACTB). Together these reagents provide positive controls for reverse transcription and SYBR™ Green-based real-time PCR, and can indicate the presence of any reverse transcription or PCR inhibitors. Real-time RT-PCR with SYBR™ ACTB Primers can also provide an endogenous control for sample normalization, as well as to confirm sufficient cell input in experiments with samples consisting of more than 100 cells per lysis.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Control TemplateXenoRNA™ control (synthetic RNA transcript)
FormatTube
Gene SymbolACTB
Green FeaturesFewer resources used and less waste, Less hazardous
No. of Reactions100 Reactions
Product LineCells-to-CT™, SYBR™
Quantity100 Reactions
Shipping ConditionDry Ice
TargetXenoRNA™ control, β-actin
Label or DyeUnconjugated
Unit SizeEach
Contents & Storage

Components:
• 100 μL Xeno™ RNA Control, 105 copies/μL
• 250 μL 20X SYBR Xeno Primers
• 250 μL 20X SYBR ACTB Primers

Amounts:
• Xeno RNA Control: approximately 100 lysis reactions
• SYBR Xeno and SYBR ACTB Primers: up to 250 x 20 μL PCRs

Storage Conditions: Store at −20°C. Do not store in a frost-free freezer.

Frequently asked questions (FAQs)

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-CT™ Control Kit FAQs