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Reference Gene Primer Sets

Validated primer sets developed for use with the microRNA LNA™ PCR primer sets. For normalization of microRNA expression levels during real-time PCR analysis.


New microRNA PCR starter kit available

We are very happy to introduce our new starter kit which gives you the chance to experience the superior performance of our microRNA qPCR system in your own hands. Find out more
  • Accurate and reliable normalization of microRNA real-time PCR data
  • Optimized primer sets for the quantification of endogenous small nucleolar or nuclear RNA molecules
  • Validated for optimal performance with the Universal cDNA synthesis kit and SYBR® Green master mix, Universal RT

Coverage

Nine reference gene primer sets are available to enable high quality data normalization and generation of reliable data from many different sample types (Table 1). These primer sets target endogenous small non-coding RNAs that are constitutively expressed in a wide variety of tissues. The primer sets have been validated as reference genes for the miRCURY LNA™ Universal RT microRNA PCR system and work optimally with the Universal cDNA synthesis kit and SYBR® Green master mix , Universal RT.

For information regarding target organisms and sequence references, please see Table 1.

MicroRNA primer sets can also be used as references. For examples, see these tech notes: Click to view information regarding microRNA LNA™ PCR primer sets


Table 1 Available reference genes
Available reference genes. (Click to learn more)

Reference genes for normalization

miRCURY LNA™ Universal RT microRNA PCR Reference genes are constitutively and relatively stably expressed across different human tissues (Figure 1). The reference genes cover a large range of Cq values offering the possibility to accurately and reliably normalize across a range of microRNA expression levels.

It is very important to always confirm the consistent and unvarying expression of a gene in all tissues and under all treatments of an experiment before choosing it as the reference. It is recommended to test a number of reference genes and use software such as GeNorm and NormFinder (found as part of the GenEx software) to choose and validate reference genes.

View experimental data describing miRCURY LNA™ Universal RT microRNA PCR

Figure 1 Expression levels of reference genes in different human tissues
Expression levels of reference genes in different human tissues. (Click to learn more)

A unique system developed specifically for microRNA profiling

The miRCURY LNA™ Universal RT microRNA PCR System offers the best available combination of performance and ease-of-use on the market because it unites two important features (Figure 2):
  • One cDNA reaction for all microRNAs – One single Universal first strand cDNA synthesis reaction is used as template, regardless of the number of microRNAs being profiled. This saves precious sample, reduces technical variation, means less pipetting and saves time in the laboratory.
  • Two LNA™-enhanced microRNA qPCR primers – Both qPCR primers are microRNA-specific and optimized with LNA™. LNA™ primers bind with high affinity, and are shorter than standard PCR primers, so that both primers can fit on the microRNA without overlapping. The result is unrivalled sensitivity and specificity, and extremely low background.

Unmatched sensitivity

Universal RT combined with LNA™-enhanced and Tm normalized primers enables accurate and reliable quantification of individual microRNAs from as little as 1 pg total RNA (Figure 3). In comparison to other microRNA real-time PCR systems using either stem-loop or standard DNA primers, the LNA™-enhanced primers offer significantly increased sensitivity, especially for AU-rich microRNAs.

Exceptional sensitivity as well as extremely low background enables accurate quantification of very low levels of microRNA without the need for pre-amplification. This makes the miRCURY LNA™ Universal RT microRNA PCR System suitable for all sample types, and especially samples with low RNA content e.g. biofluids such as serum/plasma.

Fully validated and optimized

All miRCURY LNA™ Universal RT microRNA PCR primer sets have been optimized for maximum sensitivity and thoroughly validated either by wet lab testing or in silico. In wet lab validation, over 95% of assays detect 10 microRNA copies or less in the PCR reaction (Figure 4). The primer sets have also been validated for specific amplification of the target and for minimal background signal.

Truly specific – no false positives

The incorporation of LNA™ in the qPCR primers facilitates the design of assays that can distinguish between microRNA sequences that differ by a single nucleotide. This makes it a truly specific microRNA qPCR platform – the only microRNA platform that gives no false positive signals (Figure 5). In addition, the assays can discriminate between mature microRNA sequences and precursor microRNA (Table 1).

Fast, easy and reproducible

Save time and effort in the laboratory with the 3 hour easy-to-follow protocol that minimizes pipetting. By using the same cDNA as template in all subsequent PCR reactions, the procedure is greatly simplified compared to systems that require microRNA-specific cDNA synthesis.

With fewer pipetting steps, technical variation is also reduced. As a result, it is possible to achieve extremely high reproducibility from day-to-day (Figure 6) and even site-to-site.

Flexible qPCR system

Tailor your experimental setup to your specific need using individual assays or fully flexible custom Pick-&-Mix panels . Using the same cDNA reaction, shift seamlessly between individual primers, pre-designed miRNome and Focus panels, or custom Pick-&-Mix panels.
Figure 1 Overview of the miRCURY LNA™ Universal RT PCR workflow
Overview of the miRCURY LNA™ Universal RT PCR workflow. (Click to learn more)

Figure 2 Schematic outline of the miRCURY LNA™ Universal RT microRNA PCR System
Schematic outline of the miRCURY LNA™ Universal RT microRNA PCR System. (Click to learn more)

Figure 3 Accurate quantitation from down to 1 pg total RNA starting material
Accurate quantitation from down to 1 pg total RNA starting material. (Click to learn more)

Figure 4 Serial dilution of hsa-miR-181a
Serial dilution of hsa-miR-181a. (Click to learn more)

Figure 5 Specificity results from the miRQC Study
Specificity results from the miRQC Study. (Click to learn more)

Table 1 Discrimination between mature and precursor microRNAs
Discrimination between mature and precursor microRNAs. (Click to learn more)

Figure 6 Excellent day-to-day reproducibility
Excellent day-to-day reproducibility. (Click to learn more)

Products ready to order Click to view pricing
Showing 1 - 12 of 12 products
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Prod. No.
Product Name
Description Notes
203901 SNORD38B (hsa) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203902 SNORD44 (hsa) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203903 SNORD48 (hsa) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203904 SNORD49A (hsa) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203905 SNORA66 (hsa) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203906 5S rRNA (hsa,mmu) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203907 U6 snRNA (hsa, mmu, rno) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203908 RNU5G (hsa, mmu, rno) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203909 RNU1A1 (hsa, mmu, rno) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203910 SNORD65 (mmu) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203911 SNORD68 (mmu) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]
203912 SNORD110 (mmu) PCR primer set, UniRT miRCURY LNA™ Universal RT microRNA PCR, reference gene primer set, 200 rxns [!]


NOTICE TO PURCHASER: DISCLAIMER OF LICENSE No license is conveyed with the purchase of this product under any of US Patents Nos. 5,210,015, 5,487,972, 5,804,375, 5,994,056, 6,171,785, 6,214,979, 5,538,848, 5,723,591, 5,876,930, 6,030,787, and 6,258,569, and corresponding patents outside the United States, or any other patents or patent applications, relating to the 5’ Nuclease and dsDNA-Binding Dye Processes. For further information contact the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
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