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NEW: V2.0 microRNA Ready-to-Use PCR panels

Ina K. Dahlsveen, Product manager Our V2.0 Human panels (I and II) are now available to order. V2.0 includes additional microRNA assays and improved performance. Pre-aliquoted microRNA PCR primer sets in 384-well PCR plates for exceptionally sensitive and highly specific expression profiling. Enables profiling of up to 742 human microRNAs using only 40ng total RNA – no pre-amplification necessary
Ina K. Dahlsveen, Ph.D., Product Manager
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  • Unmatched sensitivity – profile 742 microRNAs using only 40ng total RNA – no pre-amplification necessary
  • Ideal for samples with low RNA yield such as serum/plasma, FFPE sections and LCM samples
  • Fast and easy to use – just add a mix of cDNA and SYBR® Green master mix to the plates and run the real-time PCR
  • Highly reproducible – simple protocol minimizes technical variation
  • Reliable performance – fully validated and optimized LNA™-enhanced primers offer discrimination of closely related microRNAs and between mature and precursor microRNAs

Features

The microRNA Ready-to-Use PCR panels enable exceptionally sensitive high-throughput expression profiling from minimal amounts of starting material. The combination of a Universal cDNA synthesis reaction and Ready-to-Use PCR panels enables fast and easy microRNA profiling. The panels contain dried down primers for one reaction per well and simply require addition of cDNA and SYBR® Green master mix before real-time PCR amplification.

Go from sample to results in just 3 hours. See Figure 1 for an overview of the miRCURY LNA™ Universal RT microRNA PCR workflow.

The LNA™-enhanced, microRNA specific PCR primers offers 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. For more details please see microRNA LNA™ PCR primers.
 

Coverage

Two Ready-to-use PCR panels covering a total of 742 microRNAs from miRBase 13.0 are available. Human Panel I contains 375 primer sets for real-time amplification of microRNAs with high priority. These microRNAs are generally more highly expressed, most likely to be differentially expressed in disease or most often cited in the literature. Human Panel II contains 367 primer sets of microRNAs with lower priority. To view the list of primer sets in each panel, please download the plate layout files: V2.0 (new), V1.0 (old).

Both panels contain 6 reference gene primer sets , three small RNA reference genes (U6, SNORD38B and SNORD49A) and three microRNA reference genes (hsa-miR-103, hsa-miR-191 and hsa-miR-423-5p). In addition, each panel contains an inter-plate calibrator in triplicate and a control primer set for detection of a synthetic RNA spike-in provided in the Universal cDNA synthesis kit.
 

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.

Figure 1 miRCURY LNA™ Universal RT PCR workflow
Overview of the miRCURY LNA™ Universal RT PCR workflow. (Click to learn more)

Accurate microRNA expression profiling with minimal sample

The microRNA Ready-to-Use PCR panels enable profiling of 742 microRNAs or both Human panel I and II, from just 40ng total RNA. This makes it possible to do high quality microRNA expression profiling in samples that contain very little total RNA, such as FFPE sections and serum/plasma (Figures 1 and 2).

The panels contain fully validated and optimized LNA™-enhanced microRNA specific PCR primers. Please go to the microRNA LNA™ PCR primer section for details and experimental data covering primer set performance.
 

Fast, easy and reproducible

Save time and effort in the laboratory with the 3 hour easy-to-follow protocol. The synthesized cDNA is simply diluted, combined with the SYBR® Green master mix and aliquoted into all the wells of the 384-well PCR plate which is then ready to run in the real-time PCR instrument. 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 (Figure 3) and even site-to-site (Figure 4). Very high levels of reproducibility are even possible when profiling using difficult samples such as RNA purified from serum (Figure 5).
 

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.
Figure 1 Profile 730 miRNA from 40 ng total RNA
Expression profiling of 730 microRNAs using 40 ng total RNA. (Click to learn more)

Figure 2 miRNA profiling in serum
Profiling of 368 microRNAs from 35μl serum. (Click to learn more)

Figure 3 Excellent day-to-day reproducibility
Excellent day-to-day reproducibility. (Click to learn more)

Figure 4 Excellent site-to-site correlation
Excellent reproducibility between different sites. (Click to learn more)

Figure 5 miRNA qPCR reproducibility
Excellent reproducibility between RT reactions on total RNA from serum. (Click to learn more)
Note: The new V2.0 microRNA Ready-to-Use Human panels I and II will be shipping from 15 June 2010.

Important: The new V2.0 panels I and II have a different plate layout compared to V1.0 and contain improved microRNA assays.


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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