home
services
platforms
agilent: miRNA
|
The UHNMAC is now offering a miRNA expression service using the Agilent Platform. Agilent offers human, mouse, and rat miRNA arrays each containing 15,000 60-mer oligonucleotides probes generated from Sanger miRBase version 12. Probes are representative of mature miRNA species and allow for increased sensitivity due to their hairpin design. |
|
Human: 723 human miRNAs and 76 human viral miRNAs
Mouse: 567 mouse miRNAs and 10 mouse gamma herpes virus miRNAs
Rat: 350 rat miRNAs
All Agilent miRNA array format uses an "8-up" design where 8 arrays are present on each slide. Not all arrays need to be used; however, unused arrays cannot be used at a later date so pricing is based on a complete slide being used. For best pricing, experiments should be designed to use multiples of 8 arrays. For other options, please inquire.
Agilent’s miRNA Labelling Reagent and Hybridization kit use a novel Cyanine 3-nucleotide (cyanine 3-Cytidine bisphosphate (pCp)) which selectively labels and hybridises mature miRNAs. The Agilent platform is a 1-colour assay.
To learn more about Agilent's miRNA platform, please visit Agilent's miRNA webpage.
(last updated: June 2010)Please note that this pricing is for academic groups. Commercial customers should contact us for a quote.
Service includes:
- Total RNA Quality Check with Bioanalyzer & NanoDrop per sample ($15 fee is charged if RNA quality deemed too low quality to proceed with the experiment)
- Labelling of one RNA sample per array with cyanine-3
- Hybridisation/washing/scanning of arrays
- Spot quantification (Agilent Feature Extraction)
- Feature Extraction output files sent to customer (FTP)
We also offer an optional data analysis service.
Sample Requirement:
High quality total RNA is required for microarray experiments. The Agilent miRNA labelling system is a one colour system and each labelling reaction requires 100ng of total RNA.
The Agilent miRNA labelling protocol recommends total RNA; however, you must be certain to use an extraction method that does not eliminate the small RNA components. At this time miRNA enriched RNA and fractionated miRNA are not recommended.
Recommended extraction protocols include Qiagen miRNeasy, Ambion mirVanaTM, and Trizol. Extraction kits often result in the best RNA quality for this labelling protocol as they are able to more completely eliminate residual organics (such as phenols) during purification; hence, improving subsequent labelling efficiency. The 260/230 absorbance ratio is a good indicator of organic contamination and RNA of a 260/230 ratio of greater than 1.8 is required in order to proceed with the miRNA array experiment. If Trizol is the desired extraction method, additional chloroform extractions are recommended to ensure all Trizol is removed. (Please note that each additional chloroform extraction may decrease the overall RNA yield but should increase its quality).
We request a total of at least 2 µg of total RNA at a concentration of 100 ng/µL. The quality of the samples is checked upon arrival using the Agilent 2100 Bioanalyzer. An RNA Integrity Number (RIN) of 7.5 is required for guaranteed good results. Please note that if samples do not meet quality control and the customer opts not to continue with the project, a fee of $15/sample will be charged.
Array Types for miRNA profiling:
Please see the Agilent Array List for available arrays.
Custom designs are also available.
miRNA profiling is also available on the Affymetrix and Illumina array platforms.