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Exosome Focus microRNA PCR Panels

PCR panels containing miRCURY LNA™ Universal RT microRNA PCR assays for microRNAs typically found in exosomes. Panels are available in ready-to-use 96 and 384 well plates – just add cDNA and PCR master mix.

AACR Poster: A microRNA signature in urinary exosomes for diagnosis of prostate cancer

Read about Exiqon's microRNA signature in cell-free urine for diagnosis of prostate cancer, validated in independent cohorts.

Download the poster
  • All typically expressed microRNAs found in urine or cerebrospinal fluid samples in one PCR Panel
  • A highly flexible solution that allows you to focus on the microRNAs you need in your exosome project
  • Intuitive plate designer allows replacement of assays in the plates before ordering
  • Sensitivity and specificity for accurate microRNA profiling
  • Fast and easy - ready-to-use format for direct sample application with a full profile in less than 3 hours
  • High compatibility – 96 and 384 well formats compatible with most real-time PCR instruments

Why study exosomes?

Exosomes are cell derived membranous particles ranging in size from 20 to 120 nm, approximately the same size as viruses but considerably smaller than microvesicles (Figure 1). Exosomes are excreted from cells into the surrounding media and can be found in many if not all body fluids. Their proposed role as intercellular hormone-like messengers together with their stability as carrier of proteins and RNA make them ideal as biomarkers for a variety of diseases and biological processes (Figure 2).

Exosomes are secreted by most cell types and are formed by the fusion of multivesicular bodies with the plasma membrane. They are believed to be involved in a number of functions, including:
  • Immune regulation (e.g. tumor derived exosomes may help the tumor to evade the immune response)
  • Blood coagulation
  • Cell migration
  • Cell differentiation
  • Cell-to-cell communication
Microvesicles that are larger than exosomes (up to 1 µm) are typically formed by blebbing of the plasma membrane, whereas exosomes are released by exocytosis from multivesicular bodies of the endosome. 


How to isolate exosomes

The two classical ways to isolate exosomes are either by cumbersome sucrose gradient ultra-centrifugation or by isolation using antibodies specific for one of the surface proteins typically found on exosomes. The last method is very specific and might exclude some sub fractions of exosomes. The miRCURY™ Exosome Isolation Kits ensure a very high recovery of exosomes, using a straightforward and fast protocol. If used in conjunction with miRCURY™ RNA Isolation Kits, high quality microRNA can be extracted from exosomes from various biofluids. 

Although not species specific, the protocols are optimized for working with human biofluids. Recommended starting material for different organisms and sample types can be found in the manual.

Figure 1 The structure of an exosome The structure of an exosome. (Click to learn more)

Figure 2 The hypothesis of exosomal shuttling of microRNAs The hypothesis of exosomal shuttling of microRNAs. (Click to learn more)

Product coverage

The miRCURY LNA™ Universal RT microRNA PCR, Exosome Focus microRNA PCR Panels are offered as a pre-defined Pick-&-Mix microRNA PCR Panels. It is pre-defined lists of assays for microRNAs expressed in either exosomes from urine or exosomes from cerebrospinal fluid (CSF). Both assay lists are designed to fit into a single 96-well PCR plate or as 4 replicates in a 384-well PCR plate. 


The Urine Exosome Focus microRNA PCR Panels contains:
  • 87 microRNA LNA™ PCR primer sets targeting human microRNAs expressed in human exosomes from urine.
  • 5 of the microRNA assays can also be used as potential internal control primers: hsa-miR-103-3p, hsa-miR-191-5p, hsa-miR-93-5p, hsa-miR-423-3p and hsa-miR-423-5p.
  • 5 RNA spike-in controls available for selection in the Pick-&-Mix configurator for use as positive controls e.g. for quality control of the RNA purification and the RT reaction.
  • 3 inter-plate calibrators (3 x UniSp3 IPC).
Download a list of all microRNAs included on the panel (MS Excel) 


The CSF Exosome Focus microRNA PCR Panels contains:
  • 87 microRNA LNA™ PCR primer sets targeting human microRNAs expressed in human exosomes from cerebrospinal fluid.
  • 3 of the microRNA assays can also be used as potential internal control primers: hsa-miR-103-3p, hsa-miR-191-5p and hsa-miR-93-5p.
  • 5 RNA spike-in controls available for selection in the Pick-&-Mix configurator for use as positive controls e.g. for quality control of the RNA purification and the RT reaction.
  • 3 inter-plate calibrators (3 x UniSp3 IPC).
Download a list of all microRNAs included on the panel (MS Excel)


A truly flexible solution

Since the panels are offered in the Pick-&-Mix configurator as a pre-defined panels, the panel composition is not static. Assays can be added or removed according to your needs. This highly flexible solution allows you to design the plates the way you want them in 96 or 384 well formats compatible with a wide range of PCR instruments (Figure 1).


How the assays were selected

All microRNA assays in the panels were selected based on a combination of experimental data from in-house studies on exosomes, using our miRCURY LNA™ Universal RT microRNA PCR system and results published in peer-reviewed journals.

The assays are not a complete list, targeting all microRNAs found in exosomes, but they target those microRNAs typically found. If missing some microRNAs isof a concern it is recommended to run a full miRNome study using micrCURY RNA PCR Panel I+II

Figure 1
Instrument coverage of the panels. (Click to learn more)



Experimental data

Experimental data See Figure 1 for an expression profile of microRNAs from exosomes isolated from urine. Exosomes were isolated from 5 mL of urine using miRCURY Exosome Isolation Kit - Cells, urine and CSF (Prod. No 300102), followed by an RNA extraction using miRCURY RNA Isolation Kit - Cell and Plant (Prod. No 300110). The microRNA profile was examined using the Urine Exosome Focus microRNA PCR Panel.

See Figure 2 for an expression profile of microRNAs from exosomes isolated from cerebrospinal fluid. Exosomes were isolated from 1 mL of cerebrospinal fluid using miRCURY Exosome Isolation Kit - Cells, urine and CSF (Prod. No 300102), followed by an RNA extraction using miRCURY RNA Isolation Kit - Cell and Plant (Prod. No 300110). The microRNA profile was examined using the CSF Exosome Focus microRNA PCR Panel.

Figure 1
Urine microRNA profile. (Click to learn more)

Figure 2
CSF microRNA profile (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)

How to order Exosome Focus microRNA Panels

Exosome Focus microRNA Panels are ordered using our Pick-&-Mix microRNA PCR Panel plate configurator. Follow the steps below to order your plates.

1. Click on the button below. This will take you to our Pick-&-Mix microRNA PCR Panel plate configurator.


2. Register an account, sign in to your account or continue without signing in.

3. In Step 1 of the configurator, you are asked to choose between 96 and 384 well plates. Press “Next” to continue to the next step.

4. Select your PCR instrument and click “Next”.

5. In step 3 of the configurator, select the Exosome Focus panel of interest. You will be able to modify the selection of assays at a later stage. Press “Next”.

6. Please provide a name for your plate and enter your email address (email address is entered by default if you are signed in). Your email address is needed in order to send you the plate layout electronically. In this step you can also choose between different plate layouts.  For ordering the Focus Panel in a 96-well plate, you will need to select the 96 well – fully flexible layout . For ordering the Focus Panel in a 384-well plate format you can choose between the “384-well – fully flexible” layout or the “94 microRNA x 4 samples” layout.  Press “Next”.

7. In step 5 of the configurator, you are asked to choose which assays to include on the plate. Press the “Assign all pre-defined assays to panel” button to include all assays. Please note, you will need to add the four additional RNA Spike-in controls manually (UniSp2, UniSp4, UniSp5 and cel-miR-39-3p). These are to be added in the four empty wells at the bottom of the panel. You can add or remove assays from the plate. To add an assay, click the check box next to the assay in the list to the left and press the “Upload to Panel” button. To remove individual assays, click on the well in the plate depiction to select it and then click “Delete Selected Assays”.

8. If you did not log in to your account before entering the plate configurator tool, you will be asked to do so now. In step 6 of the configurator, you can save your plate configuration, make corrections or order plates by pressing “Finalize plates”. This will place the plates in your shopping basket. From here, you can either buy the plates online or request a quote from your local sales representative or distributor if you are located in a country in which Exiqon has a distributor.



NOTICE TO PURCHASER: LIMITED LICENSE Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,994,056 and 6,171,785. The purchase of this product includes a limited, nontransferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser's own internal research. No right under any other patent claim and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser's activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
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