I would like to detect specific miRNA(miR-219) in human blood, I have no idea what can I use as a positive control.
I use spike in cel-39., cel-54 and cel-238, because they have limited homologous with human mirnas. I order synthetic versions and add a known amount to a known volume of the serum/plasma at the isolation step. I tested different amounts to get it in the same approx amplification range as the mirnas I was assaying. Then when you do your pcr you also assay for the spike in MiRNA and normalize your MiRNA of interest to that. If you search MiRNA cel spike in control serum/plasma on pubmed you will find many similar papers using such methods.
Some people use a spike in control, typically something that is not found in the endogenous host.
I use spike in cel-39., cel-54 and cel-238, because they have limited homologous with human mirnas. I order synthetic versions and add a known amount to a known volume of the serum/plasma at the isolation step. I tested different amounts to get it in the same approx amplification range as the mirnas I was assaying. Then when you do your pcr you also assay for the spike in MiRNA and normalize your MiRNA of interest to that. If you search MiRNA cel spike in control serum/plasma on pubmed you will find many similar papers using such methods.
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- Biological Science
- Biochemistry
- Biomolecules
- Nucleic Acids
- RNA
- Small RNA
- microRNA