Exiqon
Home Search Contact Print
0 items in basket0
Sign In
 
 
Products Services Applications Resource Center Ordering About Exiqon
microRNA Research
lncRNA Research
mRNA Research
DNA Research
Custom Oligos
Other Reagents
RNA Isolation
Sequencing
Microarray Analysis
Real-time PCR
Northern Blotting
In Situ Hybridization
Functional Analysis
RNA Preparation
Sequencing
Microarray analysis
Real-time PCR
In Situ Hybridization
Antisense
RNA Preparation
Sequencing
Microarray analysis
Real-time PCR
In Situ Hybridization
Antisense
SNP Detection
Sequencing
Microarray Analysis
In Situ Hybridization
LNA Phosphoramidites
A2 Quencher
RNA Isolation Services
microRNA Array Services
microRNA PCR Services
Sequencing Services
microRNA ISH Services
Custom Pharma Service
Isolation
Microarray Analysis
Northern Blotting
Real-time PCR
In Situ Hybridization
Functional Analysis
SNP Detection
RNA
microRNA
RNA
DNA
microRNA
microRNA
mRNA
microRNA
Other RNAs
DNA
microRNA
RNA
DNA
microRNA Resources
mRNA and lncRNA Resources
Documents
Reading rooms
Technology
Oligo design tools
Movies
microRNA
Research Guide
Reading rooms
Application Stories
Bioinformatics and data analysis
What are lncRNAs?
Application Stories
Bioinformatics
Oligo modifications
LNA™
AQ-link
Order microRNA products
Order custom oligos
Distributors
Global Service Providers
General Information
Contact
News
History
Management
Board of Directors
Newsletter
Career
Sign up to Newsletter
Newsletter Back Issues
 

Serum / Plasma Focus microRNA PCR Panels

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

microRNA profiling in liquid biopsies

The comprehensive guide to analyzing microRNAs in liquid biopsies by qPCR or NGS.

Download Biofluids Guidelines
  • Optimal coverage – Most comprehensive platform on the market for focused microRNA profiling in serum and plasma, new v4 panel with optimized content ensures increased call-rates
  • Excellent sensitivity – Profile all microRNAs on the panels using just 20 µl serum/plasma
  • Exceptional accuracy – Provides the sensitivity and specificity required for reliable detection of microRNAs starting at just 5 microRNA copies - ideal for serum/plasma and other cell-free samples
  • Proven track record – fully experimentally validated on thousands of clinical serum/plasma samples
  • High compatibility – 96 and 384 well formats compatible with most real-time PCR instruments
  • Fast and easy – Go from samples to results in just 3 hours without the need to pre-amplify

Why study microRNAs in serum and plasma?

MicroRNAs are present in small amounts in blood serum and plasma. They circulate in the blood in a highly stable, cell-free form, suggesting that they are protected from the RNA-degrading enzymes found in blood (Figure 2).

MicroRNA profiles have been shown to be altered in serum and plasma taken from patients suffering from cancer and other diseases. This makes microRNAs promising candidates for use as a non-invasive way to discover important new biomarkers for a wide range of diseases and biological processes. Blood samples are easy to obtain and carries little risk to the patient compared to biopsies and surgical procedures.

Challenges of microRNA profiling in serum and plasma

Serum and plasma only contain a fraction of the RNA found in whole blood (Figure 3). Most of the RNA is found in the buffy coat, which mainly consists of white blood cells and platelets. In order to get accurate microRNA profiles, it is important to ensure that the serum/plasma is not contaminated by RNA from blood cells as this would skew the results of the experiment. .

There are several other challenges when it comes to profiling microRNAs in serum and plasma:
  • Small amounts of RNA means that standard ways of RNA quality control cannot be used
  • RNA from serum/plasma contains inhibitors of PCR enzymes
  • Standard reference genes, such as U6, are not present in serum and plasma, which makes choosing good reference genes difficult
In order to overcome these challenges, it is important to use an extremely accurate microRNA qPCR system. The miRCURY LNA™ Universal RT microRNA PCR system has a proven track record of being very reliable for the detection of microRNAs in serum and plasma samples.

Read our guidelines for Profiling of microRNA in Blood Serum/Plasma to learn how to successfully set up your experiment.

Figure 1
MicroRNAs circulating in blood are promising biomarkers.


Figure 2
MicroRNAs are stable in serum and plasma. (Click to learn more)

Figure 3
Serum/plasma contains small amounts of RNA. (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)

Exiqon knows about microRNA profiling in serum/plasma

Exiqon has extensive in-house knowledge of microRNA profiling in serum and plasma. We know which microRNAs are typically found in the serum/plasma of healthy individuals and which microRNAs are up or down regulated in response to disease and other biological processes (Figure 1).

All 179 miRCURY LNA™ Universal RT microRNA assays on our focus panel have been carefully selected based on our vast number of in-house analyses of microRNA expression in blood serum and plasma samples as well as on the limited number of peer-reviewed published papers available. For a complete list of microRNA assays on the panel, please refer to the plate layout file.

Over 1 million in-house and collaborative data points from serum/plasma samples collected from healthy as well as diseased individuals have been used in the selection of relevant microRNAs for the panel. This includes microRNA expression data from different disease stages from various types of cancer, neurological disorders/neurodegenerative diseases, allergies/inflammation etc. Exiqon’s database is largely based on total microRNA extracted from plasma/serum, which means that a particular circulating microRNA should be on the focus panel irrespective of whether it is preferentially protein bound, found in exosomes, micro vesicles or otherwise compartmentalized, resulting in a very comprehensive panel with broad application.

Start with minute sample sizes (20 µl serum/plasma) and get microRNA profiles for all microRNAs found in serum/plasma in just three hours, without the need to pre-amplify.

Validated for clinical use

We have taken advantage of Exiqon’s pioneering clinical diagnostic work on microRNA expression profiling in serum and plasma. This means that the panel has been thoroughly validated for use with clinical samples. Furthermore, the panel format is well-suited for a clinical workflow as it is automatable and fully compatible with standard FDA-approved qPCR equipment.


Product coverage

Exiqon’s Serum/Plasma Focus microRNA PCR Panel are available in two formats:
  • 96 well, Ready-to-Use - Two 96-well PCR plates with a total of 179 LNA™ microRNA primer sets commonly found in human serum/plasma including 7 candidate reference microRNAs. Compatible with most real-time PCR instruments (Figure 2).
  • 384 well, Ready-to-Use - One 384-well PCR plate with two replicates of 179 LNA™ microRNA primer sets commonly found in human serum/plasma including 7 candidate reference microRNAs. Compatible with most real-time PCR instruments (Figure 2).
The plates are produced in Ready-to-use format which means that the primer sets are dispensed and lyophilized in the well in an amount sufficient for one 10 µL reaction per well.


Control assays

Finding good reference genes in serum and plasma samples can be challenging. Some of the common house-keeping genes such as U6 are not present in these samples and the overall RNA concentration is low. Exiqon’s Serum/Plasma Focus microRNA PCR Panels contain 7 potential reference genes:
  • miR-103-3p, miR-191-5p and miR-423-5p. These are potential reference genes found on our miRNome panels and can be used by researchers migrating to the focus panels
  • miR-93-5p, miR-425-5p. These potential reference genes are chosen as they are usually stably expressed in serum/plasma
  • miR-451a, and miR-23a-3p can be used as a control for hemolysis. If ∆Cq(miR-23a-3p – miR-451a) is >7, it may be an indication of contamination of the serum/plasma samples.

In addition, each panels also contains qPCR assays for the 5 synthetic RNAs in the RNA Spike-in kit (cel-miR-39-3p, UniSp2, UniSp4, UniSp5 and UniSp6) that may be used for monitoring the cDNA synthesis or RNA purification (UniSP6 is also provided in the cDNA synthesis kit II). Finally, each panel contains an inter-plate calibrator in triplicate and an empty negative control.

When analyzing qPCR data, Exiqon recommends normalizing across the entire panel for best results. For more information on normalizing, please refer to our guidelines for microRNA profiling in serum/plasma.


Figure 1
Detection of all the relevant microRNAs in serum and plasma. (Click to learn more)

Figure 2
Instrument coverage of the panels. (Click to learn more)

Exiqon’s qPCR system is ideal for serum/plasma

In order to quantify the small amounts of microRNAs found in serum and plasma, a very accurate qPCR system is needed. The miRCURY LNA™ Universal RT microRNA PCR system has the sensitivity needed to accurately profile microRNAs in serum and plasma (Figure 1).

More information about the miRCURY LNA™ Universal RT microRNA PCR system

Figure 1
Exiqon's qPCR system is ideal for serum/plasma. (Click to learn more)
Products ready to order Click to view pricing
Showing 1 - 7 of 7 products
Page:
1
Prod. No.
Product Name
Description Notes
203699 Serum/Plasma Focus microRNA PCR Panel, 96 well (V4.CFX) miRCURY LNA™ Universal RT microRNA PCR, 4 sets of Ready-to-Use Serum/Plasma Focus microRNA PCR Panels. Each set of panels comprises two 96-well PCR plates with 179 LNA™ microRNA primer sets focusing on serum/plasma relevant human microRNAs, including candidate reference microRNAs and 5 Spike-in control primer sets. For use with Bio-Rad CFX96 instruments. Use with ExiLENT master mix only
203879 Serum/Plasma Focus microRNA PCR Panel, 96 well (V4.RO) miRCURY LNA™ Universal RT microRNA PCR, 4 sets of Ready-to-Use Serum/Plasma Focus microRNA PCR Panels. Each set of panels comprises two 96-well PCR plates with 179 LNA™ microRNA primer sets focusing on serum/plasma relevant human microRNAs, including candidate reference microRNAs and 5 Spike-in control primer sets. For use with Roche LightCycler 480 instruments. Use with ExiLENT master mix only
203880 Serum/Plasma Focus microRNA PCR Panel, 96 well (V4.AF) miRCURY LNA™ Universal RT microRNA PCR, 4 sets of Ready-to-Use Serum/Plasma Focus microRNA PCR Panels. Each set of panels comprises two 96-well PCR plates with 179 LNA™ microRNA primer sets focusing on serum/plasma relevant human microRNAs, including candidate reference microRNAs and 5 Spike-in control primer sets. For use with ABI Fast instrument series and ABI StepOne Plus. Use with ExiLENT master mix only
203881 Serum/Plasma Focus microRNA PCR Panel, 96 well (V4.MI) miRCURY LNA™ Universal RT microRNA PCR, 4 sets of Ready-to-Use Serum/Plasma Focus microRNA PCR Panels. Each set of panels comprises two 96-well PCR plates with 179 LNA™ microRNA primer sets focusing on serum/plasma relevant human microRNAs, including candidate reference microRNAs and 5 Spike-in control primer sets. For use with BioRad iQ series, Eppendorf, Stratagene Mx4000 and AB 7000 series instruments*. Use with ExiLENT master mix only
203882 Serum/Plasma Focus microRNA PCR Panel, 96 well (V4.ST) miRCURY LNA™ Universal RT microRNA PCR, 4 sets of Ready-to-Use Serum/Plasma Focus microRNA PCR Panels. Each set of panels comprises two 96-well PCR plates with 179 LNA™ microRNA primer sets focusing on serum/plasma relevant human microRNAs, including candidate reference microRNAs and 5 Spike-in control primer sets. For use with Stratagene Mx3000P series. Use with ExiLENT master mix only
203885 Serum/Plasma Focus microRNA PCR Panel, 384 well (V4.R) miRCURY LNA™ Universal RT microRNA PCR, 4x Ready-to-Use Serum/Plasma Focus microRNA PCR Panels supplied in two 384-well PCR plates. Each panel comprises 179 LNA™ microRNA primer sets focusing on serum/plasma relevant human microRNAs, including candidate reference microRNAs and 5 Spike-in control primer sets. For use with Roche LightCycler 480 instruments. Use with ExiLENT master mix only
203886 Serum/Plasma Focus microRNA PCR Panel, 384 well (V4.M) miRCURY LNA™ Universal RT microRNA PCR, 4x Ready-to-Use Serum/Plasma Focus microRNA PCR Panels supplied in two 384-well PCR plates. Each panel comprises 179 LNA™ microRNA primer sets focusing on serum/plasma relevant human microRNAs, including candidate reference microRNAs and 5 Spike-in control primer sets. For use with ABI 7900HT and other instruments. Use with ExiLENT master mix only


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.
  Privacy   Sitemap   Legal