Applied Biosystems™ TaqMan™ MicroRNA Reverse Transcription Kit

Catalog No.	Sufficient For
43-665-97	1000 Reactions
43-665-96	200 Reactions

Catalog No. 43-665-97 Supplier Applied Biosystems™ Supplier No. 4366597

Description

Provides the necessary components for 200 reactions of optimal performance in TaqMan™ MicroRNA Assays.

A highly specific kit that quantitates only mature miRNAs, not precursors. A sensitive kit that conserves limited samples and requires only 1 to 10 nanograms of total RNA. It is fast, simple and scalable 2-step quantitative RT-PCR assay that provides high-quality results in less than three hours.

Components of this kit are used with the RT primer provided with the TaqMan™ MicroRNA Assay to convert miRNA to cDNA.

Order Info

Shipping Conditions: Dry Ice

Specifications

• Concentration: 10X
• Content And Storage: Contains 10X RT buffer, dNTP mix w/dTTp (100 mM total), RNase inhibitor (20 U/μL), and MultiScribe™ RT enzyme (50 U/μL) for 1000 reactions. Store at -15 to -25°C.
• Detection Method: Primer-probe
• GC-Rich PCR Performance: High
• PCR Method: 2-step RT-qPCR
• Reaction Speed: Fast
• Optimal Reaction Temperature: 42°C
• Product Line: TaqMan
• Product Type: MicroRNA Reverse Transcription Kit
• Quantity: 1,000 reactions
• Reverse Transcriptase: MultiScribe™
• Shipping Condition: Dry Ice
• Sufficient For: 1000 Reactions
• For Use With (Application): miRNA analysis
• Fidelity (vs. Taq): 10 X
• Final Product Type: cDNA
• No. of Reactions: 1000 Reactions
• Reaction Format: Separate Components
• Sample Type: RNA
• Size (Final Product): 200 bp or less

Frequently Asked Questions (FAQs)

Can one use the reagents from High Capacity RT kit to make cDNA for TaqMan MicroRNA Assay?

Yes. You can use the reagents from the High Capacity cDNA Kit PN 4368814 (without RNAse Inhibitor) or 4374966 (with RNAse Inhibitor). Make sure to use the reverse transcription primer provided from the TaqMan miRNA assay for the reverse transcription.

Is it possible to combine several miRNA RT primers together for reverse transcription, and if so, how?

Yes. If you are using individual TaqMan MicroRNA Assays, you can prepare your own RT and PreAmp pools. Refer to Protocol for Creating Custom RT and Preamplification Pools using TaqMan MicroRNA Assays (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/4465407_CustomRT-preamp_UG.pdf) for detailed information. Although this protocol has been tested by our development group, we recommend that you validate the performance of the particular pool that you are interested in working with. At a minimum, we recommend running a No Template Control (NTC). The NTC is crucial to identify any primer interactions that may increase the background.

Why does the TaqMan MicroRNA Reverse Transcription Kit (4366597) protocol have an initial 16 degree C step before the reverse transcription step at 42 degrees C?

The 30 min incubation at 16°C allows the RT primer to anneal to miRNA template properly. See the complete protocol at this link.

What concentration of cDNA or range do we recommend for miRNA assays when using the TaqMan MicroRNA Reverse Transcription Kit (Cat. No. 4366596)?

1-10 ng of total RNA.

How many tubes of the MultiScribe reverse transcriptase enzyme do I get in the TaqMan MicroRNA Reverse Transcription Kit (4366597)?

1 tube containing 1 mL of MultiScribe Reverse Transcriptase at 50 U/uL. This is sufficient for 1,000 reactions.

Can I purchase the MultiScribe Reverse Transcriptase from the TaqMan® MicroRNA Reverse Transcription Kit (Cat. No 4366596) separately?

Yes, the MultiScribe Reverse Transcriptase (Cat. No. 4311235) can be purchased as a standalone product. Please see the following link.

Safety and Handling

WARNING: Cancer and Reproductive Harm - www.P65Warnings.ca.gov

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