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Applied Biosystems™ High-Capacity RNA-to-cDNA™ Kit

Catalog No. 4387406
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4387406 50 Reactions
4388950 500 Reactions
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Catalog No. 4387406 Supplier Applied Biosystems™ Supplier No. 4387406
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Description

Includes

50 Reaction Kit: 1 x 50μL of 20X Enzyme mix (MuLV and RNase inhibitor protein); 1 x 500μL of 2X RT Buffer Mix (includes dNTPs, random octamers, and oligo dT-16)

The High Capacity RNA-to-cDNA Kit is a streamlined reverse transcription kit designed for optimum performance with TaqMan Gene Expression Master Mix, Power SYBR Green PCR Master Mix, and other PCR enzymes.

The High Capacity RNA-to-cDNA Kit is a streamlined reverse transcription kit designed for optimum performance with TaqMan Gene Expression Master Mix, Power SYBR Green PCR Master Mix, and other PCR enzymes. The components of the two-tube kit work together to provide sensitive and specific reverse transcription across a broad range of template amounts.

Features of the High Capacity RNA-to-cDNA Kit include:
• Fast reaction time (typically 30–60 min)
• Convenient and simple workflow with few pipetting steps
• Reliable reverse transcription of both abundant and limited targets
• Optimized two-step protocol enables multiple PCR reactions from a single RT reaction

Streamline cDNA synthesis while maintaining detection sensitivity
Amplification of a dilution series of the RNA concentration reference standard demonstrates exceptional reverse transcription and PCR efficiency across 11 orders of magnitude. This wide dynamic range allows one set of reaction conditions to detect transcripts from highly active as well as weakly expressed genes. The two-tube formulation reduces reaction time and requires fewer pipetting steps than with the High Capacity cDNA Reverse Transcription Kit, while maintaining high performance.

Order Info

Shipping Condition: Dry ice

Specifications

Content And Storage

• 20X Enzyme mix (1 x 50 μL)
• 2X RT Buffer Mix (1 x 500 μL)

Store at –15 to –25°C.
Buffer Mix may be stored at 2–8°C for up to 6 months.
Format Kit
Reaction Speed 30 to 60 min.
Technique Reverse Transcription
Optimal Reaction Temperature 37°C
Quantity 50 reactions
Reverse Transcriptase MultiScribe
Ribonuclease H Activity Yes
Shipping Condition Dry Ice
For Use With (Application) Real Time PCR (qPCR), RT-PCR
Final Product Type First-Strand cDNA
No. of Reactions 50 Reactions
Reaction Format Premixed Components
Reagent Type Reverse Transcription
Size (Final Product) Up to 7 kb
Starting Material RNA
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Frequently Asked Questions (FAQs)

How long can I store the cDNA from my reverse transcription step?

You can store your cDNA at 2-6 degrees C for up to 24 hours. For long-term storage, store the cDNA at -15 to -25 degrees C and add EDTA to a final concentration of 1 mM to prevent degradation.

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.
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.
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).
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).
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.
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.

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.

For Research Use Only. Not for use in diagnostic procedures.