New RT-qPCR users can take advantage of TAmiRNA´s osteomiR service.
Technical notes provide our users with additional information about usage, handling and technical performance of our kits. This list of technical notes will be expanded in regular intervals.
TN01 - The impact of sample type on osteomiR detection - download tech note (Oct 2016)
TN02 - Analysis of data generated using the osteomiR™ kit - download tech note (Dec 2016)
1. How many samples can we measure with one osteomiR™ kit?
One kit provides sufficient reagents for the analysis of 48 serum or plasma samples. Upon request, we can provide smaller kits for proof-of-concept studies involving 10 samples or less.
2. Can we use rodent or other non-human samples for the kits?
The osteomiR™ kit is intended to be used with human samples. However, orthologs for osteomiRs do exist in other species, allowing the use of adapted version of the tests in non-mammalian species. To assess the feasibility of such projects, please contact us before using the assay in non-human species.
3. Correlations of osteomiRs to other bone Parameters?
We have performed an in-depth analysis of the correlations of osteomiRs to parameters of dynamic histomorphometry and nanoCT. The results from this analysis have been presented during ECTS 2017 and have been submitted for publication.
4. Does the osteomiR™ kit require a specific instrument or can it be run with any qPCR instrument?
We recommend using the Roche LightCycler® 480. Other qPCR instruments can be used with the kit, but the software currently supports only data generated with Roche instruments. Data generated with ABI or other platforms can be analyzed manually. We will provide support in such instances.
5. Which quality controls are included in the assay? Should I include additional controls?
5 control primers to monitor assay and sample quality are included in the kit, no further controls are necessary.
6. I want to analyze a sample type that is not listed as tested yet, can I do that?
Please contact us to discuss the possibilities.
7. What is the recommended RNA extraction method?
All our kits include a protocol and reagents for RNA extraction. TAmiRNA´s osteomiR™ kit has been standardized to a volume of 200 µL serum as starting material to ensure high RNA yield and prevent inhibition of downstream PCR amplification using the miRCURY™ RNA isolation kit, which is included in the osteomiR™ kit. For further information read our guidelines detailing RNA purification and sample preparation for your osteomiR™ kit.
8. What is the sensitivity of the Assay?
LOD: 10 copies/µl (6 replicates, 95% positive detection); LLOQ: 35 copies/µl
9. What is the reproducibility of the assay?
Intra-assay (n=6) ≤ 20%, Inter-assay (n=3) ≤ 20%
10. What data analysis tools do you recommend?
We recommend using the software included in the kit for normalization and quality control.