Can I use the Cells-to-CT products with organoids?
Yes. For organoid applications, we recommend the Cells-to-CT Fast Advanced Kit. This kit has been evaluated for direct gene expression analysis from organoid samples and is supported by application data demonstrating its performance.
Can the TaqMan Cells-to-CT Express 1-Step Kit be integrated into automated or high-throughput workflows?
Yes. The TaqMan Cells-to-CT Express 1-Step Kit can be integrated into automated and semi-automated RT-qPCR workflows because it eliminates RNA purification and reduces sample preparation steps. For best results, assembled RT-qPCR reactions should be run promptly after setup.
How long does it take to generate RT-qPCR results using the TaqMan Cells-to-CT Express 1-Step Kit?
Researchers can move from cultured cells to RT-qPCR results in less than 90 minutes using the TaqMan Cells-to-CT Express 1-Step Kit. The workflow includes approximately 5 minutes for cell lysis followed by a single RT-qPCR reaction.
How does the TaqMan Cells-to-CT Express 1-Step Kit compare to RNA extraction-based RT-qPCR workflows?
The TaqMan Cells-to-CT Express 1-Step Kit enables RT-qPCR directly from cultured cell lysates without RNA purification. In internal evaluations, the TaqMan Cells-to-CT Express 1-Step Kit produced Ct values comparable to those obtained using RNA purified with spin column-based extraction methods while reducing sample preparation steps and workflow time.
How many freeze-thaw cycles can the Express Lysis Solution tolerate?
The Express Lysis Solution included in the TaqMan Cells-to-CT Express 1-Step Kit has been tested through at least five freeze-thaw cycles without loss of performance. For laboratories that use the reagent frequently, the Express Lysis Solution can be stored at room temperature or 2-8 degrees C for up to one year.
Can I store the Express Lysis Solution included in the TaqMan Cells-to-CT Express 1-Step Kit at room temperature?
Yes. The Express Lysis Solution can be stored at room temperature or at 2-8 degrees C after thawing. The TaqMan Cells-to-CT Express 1-Step Kit is shipped frozen and may be stored frozen until first use. Follow the storage recommendations provided in the User Guide for all kit components.
Can I freeze and store lysates prepared using the TaqMan Cells-to-CT Express 1-Step Kit?
Yes. Lysates prepared using the TaqMan Cells-to-CT Express 1-Step Kit are stable when stored at -80 degrees C for at least one year. Lysates have also been shown to tolerate up to five freeze-thaw cycles without a measurable loss in RT-qPCR signal.
Can the TaqMan Cells-to-CT Express 1-Step Kit be used with TaqMan Array Cards and Plates?
Yes. The TaqMan Cells-to-CT Express 1-Step Kit is compatible with TaqMan Array Cards and TaqMan Array Plates. When using these formats, follow the recommended sample input volumes and setup guidelines provided in the applicable TaqMan Array protocols.
Is the TaqMan Cells-to-CT Express 1-Step Kit compatible with TaqMan Gene Expression Assays?
Yes. The TaqMan Cells-to-CT Express 1-Step Kit is designed for use with TaqMan Gene Expression Assays and has been evaluated with a wide range of assay targets. For new targets or cell types, we recommend confirming performance using the pilot experiment described in the User Guide.
Can the TaqMan Cells-to-CT Express 1-Step Kit be used for multiplexed qPCR?
Yes. The master mix included in the TaqMan Cells-to-CT Express 1-Step Kit supports multiplex RT-qPCR and has been evaluated for use in reactions containing up to three probes (triplex assays). Because multiplex performance depends on assay design and target abundance, users should validate their specific assay combinations.
Does the TaqMan Cells-to-CT Express 1-Step Kit contain RNase inhibitor?
Yes. The Express Lysis Solution included in the TaqMan Cells-to-CT Express 1-Step Kit contains components that inhibit RNase activity during cell lysis, helping preserve RNA for downstream RT-qPCR analysis.
Does the TaqMan Cells-to-CT Express 1-Step Kit contain reagents for genomic DNA (gDNA) removal?
Yes. The TaqMan Cells-to-CT Express 1-Step Kit includes Express ezDNase, a double-stranded DNA-specific, heat-labile DNase that can be added during cell lysis to remove genomic DNA. Express ezDNase is automatically inactivated during the RT-qPCR thermal cycling protocol. For additional protection against gDNA signal, we recommend using TaqMan Gene Expression Assays designed to span exon-exon junctions or intron-exon boundaries, such as assays with an "_m1" suffix.
Will the TaqMan Cells-to-CT Express 1-Step Kit work for my gene target of interest?
The TaqMan Cells-to-CT Express 1-Step Kit has been evaluated with a broad range of TaqMan Gene Expression Assays and has demonstrated performance comparable to workflows using purified RNA. Because assay performance can vary depending on the target and cell type, we recommend performing the pilot experiment described in the User Guide when evaluating a new assay.
What is the maximum amount of lysate that I can use in the RT-qPCR reaction using the TaqMan Cells-to-CT Express 1-Step Kit?
Up to 10% of the total RT-qPCR reaction volume can consist of lysate prepared using the TaqMan Cells-to-CT Express 1-Step Kit. For example, up to 2 µL of lysate can be added to a 20 µL RT-qPCR reaction. Exceeding the recommended lysate input may affect RT-qPCR performance.
What cell types are compatible with the TaqMan Cells-to-CT Express 1-Step Kit?
The TaqMan Cells-to-CT Express 1-Step Kit is designed for gene expression analysis from cultured mammalian cells. Researchers should consult the product documentation for specific recommendations regarding cell lines, assay setup, and workflow optimization. To confirm compatibility with your specific cell type, follow the Pilot Experiment outlined in the User Guide.