TaqMan™ MicroRNA Reverse Transcription Kit, 1,000 reactions - FAQs

View additional product information for TaqMan™ MicroRNA Reverse Transcription Kit - FAQs (4366597, 4366596)

7 product FAQs found

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 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.

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

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.

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

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.

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

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.

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