miRNA Analysis and Non-Coding RNA Support—Getting Started

Yes. For additional support, please go here.

Yes. For additional support, please go here.

The TaqMan™ MicroRNA Assay employs a novel target-specific stem-loop reverse transcription primer to address the challenge of the short length of mature miRNA. The primer extends the 3' end of the target to produce a template that can be used in standard TaqMan™ Assay-based real-time PCR. Also, the stem-loop structure in the tail of the primer confers a key advantage to this assay: specific detection of the mature, biologically active miRNA; it does not detect the precursor. For more information, please go here.

The TaqMan™ Advanced miRNA workflow has a universal RT and also includes a universal amplification step (miR-Amp) that is built into the workflow. The first generation TaqMan™ MicroRNA workflow uses miRNA-specific stem loop RT primers and has an optional pre-amplification step that is not built into the workflow and is sold separately. For more information, please go here.

miR-Amp is a 30-minute universal amplification that uses 14 cycles of PCR to provide a 3-10 C_t improvement in sensitivity (depending on sample type). mir-Amp uses universal primers that recognize regions of the cDNA outside of the miRNA target sequence. These sequences are added onto all mature miRNAs in the sample during the cDNA synthesis workflow. The miR-Amp step does not introduce bias.

It is possible to find more than one TaqMan™ Advanced miRNA assay for a particular miRNA target. However, keep in mind that these assays are not exactly the same. Please check the column for ‘Assay Design’, and pick the one for your desired species. When you do this, there should only be one assay to choose from. The reason this occurs is that while the miRNA sequences may be conserved among certain species, our designs are cross checked against the species background of the intended species only (i.e., the human assay is checked within human genome only). So for example, assay ID 478224_mir, for hsa-mir-100-5p, lists Species: Human, Mouse, and Rat and Assay Design: Human. This assay is therefore meant for use with human samples. If used with mouse or rat samples, there may be some cross-reactivity with other isomiRs in those species.

The TaqMan™ Advanced miRNA Assay does not detect snRNAs or snoRNAs. Do not use snRNAs and snoRNAs as endogenous controls for this assay, as they do not have the 5’-phosphate that is required for the chemistry of the TaqMan™ Advanced miRNA assays.

You can easily see all the content on the microRNA assays by downloading this list. You can then use the column headers to the right to sort by either human or rodent for the respective array content.

Yes. You can use the TaqMan™ MicroRNA Assay for absolute quantitation when you need to know the exact number of copies being expressed. For this type of analysis, you will need to obtain a synthetic microRNA to create a standard curve. You can order a synthetic RNA oligo for this purpose from here.

We do have a positive control siRNA targeting MALAT-1, which is a ncRNA localized mostly in the nucleus (Cat. No. 4455877). Silencer™ Select Negative Controls No. 1 (Cat. Nos. 4390843, 4390844 and No. 2 (Cat. Nos. 4390846, 4390847) can be used as the negative controls.