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 in 8x15K or SurePrint G3 8x60K formats containing 60-mer oligonucleotides probes generated from different
Sanger miRBase versions. Probes are representative of mature miRNA species and allow for increased sensitivity due to
their hairpin design.
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,
- SurePrint G3 8x60K v16: 1205 human miRNAs and 144 human viral miRNAs (miRBase 16.0)
- 8x15K v3: 851 human miRNAs and 88 human viral miRNAs (miRBase 14.0)
- 8x15K v2: 723 human miRNAs and 76 human viral miRNAs (miRBase 10.1)
- SurePrint G3 8x60K v17: 1079 mouse miRNAs and 57 mouse viral miRNAs (miRBase 17.0)
- SurePrint G3 8x60K v16: 1023 mouse miRNAs and 57 mouse viral miRNAs (miRBase 16.0)
- 8x15K v2: 627 mouse miRNAs and 39 mouse viral miRNAs (miRBase 12.0)
- 8x15K v16: 677 rat miRNAs (miRBase 16.0)
- SurePrint G3 8x60K: Up to 60K probes per array in 8x60K format
- 8x15K: Up to 15K probes per array in 8x15K format
To learn more about Agilent's miRNA platform, please visit Agilent's miRNA webpage.
- 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. Please
enquire directly for more information.
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 mirVana, 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.