Cells-to-CT™ Bulk Lysis Reagents
Cells-to-C<sub>T</sub>&trade; Bulk Lysis Reagents
Citations & References (3)
Invitrogen™

Cells-to-CT™ Bulk Lysis Reagents

Green features
Cells-to-CT™ Bulk Lysis Reagents are components of the TaqMan™ Gene Expression Cells-to-CT™ Kit, here made available as a separate purchase,Read more
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Catalog NumberQuantity
4391851C2,500 reactions
Catalog number 4391851C
Price (CLP)
-
Quantity:
2,500 reactions
Cells-to-CT™ Bulk Lysis Reagents are components of the TaqMan™ Gene Expression Cells-to-CT™ Kit, here made available as a separate purchase, in larger volumes than provided with the kit. The reagents included are: Lysis Solution, Stop Solution, and DNase. The TaqMan™ Gene Expression Cells-to-CT™ Kit enables researchers to easily perform expression analysis using cultured cells without purification of RNA.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Green FeaturesFewer resources used and less waste, Less hazardous
No. of Reactions2500 reactions
Product LineCells-to-CT
Product TypeCell Lysis Buffer
Quantity2,500 reactions
Shipping ConditionDry Ice
Unit SizeEach
Contents & Storage

This kit contains the following:
• Lysis Solution (124 mL): Store at 4°C
• Stop Solution (12.5 mL): Store at -20°C
• DNase (1.25 mL): Store at -20°C

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 Cells-to-CT™ Bulk Lysis Reagents FAQs

Citations & References (3)

Citations & References
Abstract
Genome-wide siRNA-based functional genomics of pigmentation identifies novel genes and pathways that impact melanogenesis in human cells.
Authors:Ganesan AK, Ho H, Bodemann B, Petersen S, Aruri J, Koshy S, Richardson Z, Le LQ, Krasieva T, Roth MG, Farmer P, White MA,
Journal:PLoS Genet
PubMed ID:19057677
'Melanin protects the skin and eyes from the harmful effects of UV irradiation, protects neural cells from toxic insults, and is required for sound conduction in the inner ear. Aberrant regulation of melanogenesis underlies skin disorders (melasma and vitiligo), neurologic disorders (Parkinson''s disease), auditory disorders (Waardenburg''s syndrome), and opthalmologic disorders ... More
N-methylpurine DNA glycosylase and DNA polymerase beta modulate BER inhibitor potentiation of glioma cells to temozolomide.
Authors:Tang JB, Svilar D, Trivedi RN, Wang XH, Goellner EM, Moore B, Hamilton RL, Banze LA, Brown AR, Sobol RW,
Journal:Neuro Oncol
PubMed ID:21377995
'Temozolomide (TMZ) is the preferred chemotherapeutic agent in the treatment of glioma following surgical resection and/or radiation. Resistance to TMZ is attributed to efficient repair and/or tolerance of TMZ-induced DNA lesions. The majority of the TMZ-induced DNA base adducts are repaired by the base excision repair (BER) pathway and therefore ... More
Accurate RT-qPCR gene expression analysis on cell culture lysates.
Authors:Van Peer G, Mestdagh P, Vandesompele J,
Journal:Sci Rep
PubMed ID:22355736
'Gene expression quantification on cultured cells using the reverse transcription quantitative polymerase chain reaction (RT-qPCR) typically involves an RNA purification step that limits sample processing throughput and precludes parallel analysis of large numbers of samples. An approach in which cDNA synthesis is carried out on crude cell lysates instead of ... More