Download biomedica product list 2018osteomiR™ – miRNA Biomarkers
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Assay time
Sample to data in 8 hours with 2 hours hands-on time
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Sample volume
200 µl
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Sample type
Serum, EDTA plasma
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Method
RT-qPCR, relative quantification without standard curve
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Sensitivity
LOD: 10 copies/µl (6 replicates, 95% positive detection); LLOQ: 35 copies/µl
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Regulatory status
Research use only.
Product Overview
We have identified 19 microRNAs in human serum which
» are associated to bone microstructure and histomorphometry
» are regulated in patients with osteoporotic fractures
» are BMD-independent risk factors for osteoporotic fractures
» regulate bone formation and resorption via multiple pathways
» are novel biomarkers for bone diseases
» can easily be detected in human serum and plasma samples.
Principle Of The Osteomir™
A universal reverse transcription (RT) reaction is performed, as shown in the Figure below (Step 1), which means that all microRNA species are converted into complementary DNA (cDNA) at the same time. This enables parallel quantitative PCR (qPCR) detection of different microRNA sequences in one cDNA sample using the osteomiR™ plate. Universal RT is achieved by first adding a poly-A tail to the mature microRNA template (Step 1). Complementary cDNA is synthesized using a poly-T-primer with a 3’ degenerate anchor and a 5’ universal tag. During qPCR, the cDNA is then amplified using microRNA-specific and LNATM-enhanced forward and reverse primers using SYBR® Green for detection of double-stranded DNA (Step 2).
Kit Components
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Contents |
Description |
Quantity |
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qPCR plates |
Storage: -20°C; primer coated 96 or 384 well qPCR plates, 19 osteomiRs™ and 5 controls/sample per plate |
48 samples |
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Serum/Plasma RNA extraction Kit |
Storage: RT |
48 samples |
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osteomiR chemistry |
Storage: -20°C; Spike-Ins, RT chemistry, SYBR Green Mix |
48 samples |
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SOFTWARE |
osteomiR™ software included to obtain normalized data and a fracture-risk score |
Serum Collection
The osteomiR™ Workflow requires 200 μL serum.
Serum collection is ideally performed after overnight fasting in the morning hours between 8 am and 10 am. This can reduce biological variance in miRNA levels due to activity and diet. We recommend the use of 21 gauge needles and red top vacutainer tubes (BD vacutainer®) for blood collection. Filled collection tubes should sit upright after the blood is drawn at room temperature for a minimum of 30 to a maximum of 60 minutes for the clot to form. The red top tubes do not have to be full to be used.
Centrifuge the blood sample at 2 500 g for 10 minutes at room temperature in a horizontal rotor (swing-out head). If the blood is not centrifuged immediately after the clotting time (30 to 60 minutes at room temperature), the tubes should be refrigerated (4ºC) for no longer than 4 hours. After centrifuging, the clot is located at the bottom of the tube, and the serum is on top of the clot. Use a clean pipette and nuclease-free filter tips to transfer the serum (recommendation: do not pour!). Pipette serum into the labeled nuclease-free (1.5 ml) tubes, filling the vials in sequential order. Aliquot volume is recommended to be 225 μL, so that 200 μL can be safely used for RNA extraction. Close the caps on the vials tightly. This process should be completed within 1 hour of centrifugation.
Check that all aliquot vial caps are secure and that all vials are labeled. Place all aliquots upright in a specimen box or rack in an -80ºC or colder freezer. All specimens should remain at -80ºC or colder prior to analysis or shipping. The sample aliquots should not be thawed prior to analysis or shipping.
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Bone Diseases
- Bone turnover
- Microstructure & histomorphometry
- Osteoporosis & other bone diseases
- Bone loss & treatment response
- Calcification
- Therapeutic activity
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Literature
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Changes of Circulating MicroRNAs in Response to Treatment With Teriparatide or Denosumab in Postmenopausal Osteoporosis.
Anastasilakis AD. et al.,
J Clin Endocrinol Metab, 2018; 1;103(3):1206-1213.
PMID:29309589 -
Bone-related Circulating MicroRNAs miR-29b-3p, miR-550a-3p, and miR-324-3p and their Association to Bone Microstructure and Histomorphometry.
Feichtinger X. et al.,
Sci. Rep., 2018; 20;8(1):4867.
PMID:29559644 -
Circulating microRNA signatures in patients with idiopathic and postmenopausal osteoporosis and fragility fractures.
Kocijan R, et al.,
J Clin Endocrinol Metab., 2016; 101(11):4125-4134.
PMID:27552543 -
Serum microRNAs are indicative of skeletal fractures in postmenopausal women with and without type-2 diabetes and influence osteogenic and adipogenic differentiation of adipose-tissue derived mesenchymal stem cells in vitro.
Heilmeier U. et al.,
J Bone Miner Res., 2016; 31(12):2173-2192
PMID:27345526
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PMID:29309589