microRNA LNA™ PCR primer sets

A unique system for microRNA profiling

The miRCURY LNA™ Universal RT microRNA PCR offers the best available combination of performance and ease-of-use on the microRNA real-time PCR market because it unites two important features (Figure 1):

• Universal RT – One first-strand cDNA synthesis reaction (or RT reaction) can be used as template for multiple microRNA real-time PCR assays. This saves precious sample, reduces technical variation and saves time in the laboratory.
• LNA™ PCR amplification – Both PCR amplification primers (forward and reverse) are microRNA specific and optimized with LNA™. The result is 1) exceptional sensitivity as well as extremely low background enabling accurate quantitation of very low microRNA levels and 2) highly specific assays that allow discrimination between closely related microRNA sequences.

Coverage

A total of 1393 wet-lab validated LNA™ enhanced PCR primer sets are available covering human, mouse, rat and viral microRNAs. For complete details of coverage please download the complete assays list using the link below. To find a particular primer set, please use the search function below. All miRCURY LNA™ Universal RT microRNA PCR primer sets have been optimized and strictly validated for specific amplification of the target and for minimal background signal. If your microRNA of interest is not covered by the standard offering below, custom designed LNA™ microRNA primer sets for any microRNA are available.
 

Note:

The microRNA LNA™ PCR primers have been optimized for use with the Universal cDNA synthesis kit and ExiLENT SYBR® Green master mix kit. Use of other reagents will affect the quality of the results.

Click here for information on Reference gene primer sets.

Unmatched sensitivity

The exceptional sensitivity of the microRNA PCR primer sets is achieved by combining the following features:

• Universal RT
• LNA™ enhanced and Tm normalized primers

This combination enables accurate and reliable quantification of individual microRNAs from as little as 1 pg of total RNA input in the initial first-strand cDNA synthesis (Figure 1).

In comparison to other microRNA real-time PCR systems using either stem-loop (Figure 2) or standard (Figure 3) DNA primers, the LNA™-enhanced primers offer significantly increased sensitivity, especially for AT rich microRNAs.

The low sample requirements also enables microRNA quantitation using total RNA purified from difficult samples such as LCM samples and serum/plasma (Figure 4 and 5).

Fully validated and optimized

All miRCURY LNA™ Universal RT microRNA PCR primer sets have been optimized to be as sensitive as possible. Over 80% of assays detect minimum 100 microRNA copies in the PCR reaction whereas close to 50% detect as little as 10 microRNA copies (Figure 6). The primer sets have also been validated for specific amplification of the target and for minimal background signal (Figure 7).

Excellent specificity

The incorporation of LNA™ in the PCR amplification primers (forward and reverse) facilitates the design of assays that can distinguish between microRNA sequences that differ by only one nucleotide (Table 1). In addition, the assays can discriminate between mature microRNA sequences and precursor microRNA (Table 2).
 

Fast, easy and reproducible

Save time and effort in the laboratory with the 3 hour easy-to-follow protocol. By using the same RT reaction as template in all subsequent PCR reactions, the procedure is greatly simplified compared to systems that require microRNA-specific first-strand synthesis. The number of pipetting steps is reduced to a minimum and technical variation is minimized. As a result, it is possible to achieve extremely high reproducibility from day-to-day and even site-to-site.

Note:

The microRNA Ready-to-use PCR panels have been optimized for use with the Universal cDNA synthesis kit and SYBR Green master mix kit. Use of other reagents will affect the quality of the results.