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High-Capacity RNA-to-cDNA™ Kit, 50 reactions - FAQs

High-Capacity RNA-to-cDNA™ Kit, 50 reactions - FAQs

View additional product information for High-Capacity RNA-to-cDNA™ Kit - FAQs (4387406, 4388950)

9 product FAQs found

Can I order the RNase Inhibitor from the High-Capacity cDNA Reverse Transcription Kit separately?

Yes. You can order the RNase Inhibitor separately as Cat. No. N8080119.

What type of primers are in the High-Capacity RNA-to-cDNA Kit?

The High Capacity RNA-to-cDNA Kit includes a 2X RT buffer mix, which is composed of dNTPs, random octamers, and oligo(dT)16.

When using the PowerUp SYBR Green Master Mix (Cat. No. A25741, A25742, A25743, A25776, A25777, A25778, A25779, A25780, A25919), I get high Ct values. What can I do?

It could be due to the following reasons:
  • Insufficient cDNA template is present. Use 10 to 100 ng of cDNA template per 50 µL reaction.
  • Quality of cDNA template is poor. Quantify the amount of cDNA template and test the cDNA template for the presence of PCR inhibitors.
  • Sample degradation. Prepare fresh cDNA, then repeat the experiment.
  • Reduced number of PCR cycles in the thermal cycling protocol. Increase the number of PCR cycles to the default setting of 40.
  • Primer-dimer formation and residual polymerase activity. Prepare the reaction mixes and reaction plate on ice. To ensure optimal results, run the reaction plate as soon as possible after completing the reaction setup.


Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

When using the PowerUp SYBR Green Master Mix (Cat. No. A25741, A25742, A25743, A25776, A25777, A25778, A25779, A25780, A25919), the Multicomponent signal for ROX is not linear. What could cause this?

This could be due to the following reasons:
  • Pure dye component’s spectra are incorrect. Rerun pure dye spectra.
  • Incorrect dye components have been chosen. Choose correct dyes for data analysis.


Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

Which reverse transcription kit can I use with the PowerUp SYBR Green Master Mix (Cat. No. A25741, A25742, A25743, A25776, A25777, A25778, A25779, A25780, A25919)?

We recommend using the High-Capacity RNA-to-cDNA Kit (Cat. No. 4387406, 4388950).

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

Which kit should I use for the reverse transcription step for TaqMan Pri-miRNA assays (Cat. No. 4427012)?

We recommend the following kits:
  • High-Capacity cDNA Reverse Transcription Kit (Cat. No. 4374966).
  • High-Capacity RNA-to-cDNA Kit (Cat. No. 4387406).


Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Is the cDNA generated using the High-Capacity RNA-to-cDNA Kit (Cat. No. 4387406, 4388950) single-stranded or double-stranded?

The cDNA generated using the High-Capacity RNA-to-cDNA Kit (Cat. No. 4387406, 4388950) is single-stranded.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Is the 2X RT Buffer Mix within the High-Capacity RNA-to-cDNA Kit (Cat. N. 4387406, 4388950) available separately?

No, the 2X RT Buffer Mix is not available separately.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

Can cDNA be generated using the High Capacity cDNA Reverse Transcription kit (SKU 4322171) for quantitative qPCR applications?

Yes, the single-stranded cDNA generated via the High-Capacity cDNA Reverse Transcription can be used in real-time PCR applications. In addition, we also recommend Superscript VILO cDNA Synthesis Kit, High Capacity RNA to cDNA kit, and/or High Capacity RNA to cDNA Master Mix for total RNA.