The unique priming of the EPIK™ miRNA Assays confers distinct advantages over other approaches to miRNA detection. EPIK miRNA Assays use modified stem-loop, miRNA-specific oligonucleotide mediated reverse transcription and hemi-nested real-time PCR, combined with SensiSMART™ and SYBR® Green, offering remarkable sensitivities as well as extremely low background, thereby enabling the accurate detection of very low input miRNA levels and accurate discrimination of even very closely related miRNA targets.
- Increased sensitivity RT and qPCR steps optimized to drive highly efficient amplification from limiting amounts (≥10 pg) of total RNA
- Improved specificity pre-designed, novel, miRNA-specific primers for superior discrimination of even very closely related targets
- Faster protocol RNA to Ct in less than 2 hours for earlier results and increased throughput capacity
- Reliable data 7-log linear dynamic range for accurate quantification of low- and high- expressed targets in even limited sample volumes
- Convenience all necessary components included in the kit and formulated to minimize set-up time
Using advanced design concepts, a proprietary algorithm has been developed by MiRXES™ to create modified stem-loop miRNA specific reverse transcription primers and hemi-nested real-time PCR primer combinations, to maximize miRNA detection sensitivity while minimizing non-specific interactions. The assays use SYBR® Green rather than a probe-based system for detection, allowing rapid amplification when required (Fig. 1). The resulting real-time PCR assays use SensiSMART™ enabling detection of extremely low levels of miRNA with high specificity, allowing the discrimination between closely related miRNA sequences (Fig. 2).
Performance of these assays have been validated to detect as few as 100 copies of template per RT reaction (Fig. 3) with excellent assay efficiency and linearity, to produce complete systems for miRNA profiling of biofluids.
The EPIK Biofluid miRNA Panel Assay is made up of the most commonly studied 176 miRNAs in circulating biofluids (see plates). A set of controls present on this assay along with spike RNA enables data analysis, assessment of reverse transcription performance and assessment of PCR performance. Circulating miRNAs are regarded as a promising new class of biomarkers and a novel means of extra-cellular communications.
EPIK miRNA Panel Assays are intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.
If you need single assays, please contact Bioline Technical Support for advice.
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Instrument CompatibilityPlate 0.1Y = ABI Fast 96-Well Block (0.1 mL, low profile) which is compatible with ABI Step One Plus.
Certification of Analysis (COAs)
|Biofluid Assay Plates||2 x 2 96 well plates|
|RT Primer Pool - lyophilized||2 tubes (A and B)|
|RNA Spike - lyophilized||1 tube|
|EPIK RT Buffer||32 µL|
|EPIK RT Enzyme||8 µL|
|2 x SensiSMART™ Master Mix||4 x 1 mL|
|DEPC Water||3 x 1.8 mL|
Storage & Stability
All components are shipped on dry/blue ice and should be stored at -20°C upon receipt for optimum stability and the RNA Spike stored at -80°C after reconstitution. Repeated freeze/thaw cycles should be avoided. When stored under the recommended conditions and handled correctly, full activity of the reagents is retained until the expiry date on the outer box label.
On Dry Ice or Blue Ice.
Yes, however all the cDNA reactions must be treated identically, so if it is not possible to run all the plates within one day, all the cDNA reactions must be frozen at ‐20°C once created. This will ensure that all cDNA reactions are subjected to the same number of freeze-thaw cycles.
0.1Y ABI 96-Well Block (0.1 mL, low profile), 0.1X ABI 96-Well Block (0.1 mL) and 0.2Y ABI 96-Well Block (0.2 mL), you need to choose the correct plate type for your instrument, this can be done using the 'Choose the Right Plate' button on each of the panel pages.