Consider we have any sample taken from patient. What technique can be used to identify specific miRNA present in that sample?
Hi
You should first do total RNA or microRNA isolation then depend on your disease of interest you can do different method like:
1- read some paper related to particular disease and find which miRNA is related then run a RT-PCR to detect the miRNA and compare with normal sample.
2-you can use prediction software and predict miRNAs related to gene then use RT-PCR to check expression.
3-you can use micro-array or PCR-array to find different expression pattern in patient and healthy person,also you can do deep seq or RNA-seq.
Best,
Mohsen
Hi
You should first do total RNA or microRNA isolation then depend on your disease of interest you can do different method like:
1- read some paper related to particular disease and find which miRNA is related then run a RT-PCR to detect the miRNA and compare with normal sample.
2-you can use prediction software and predict miRNAs related to gene then use RT-PCR to check expression.
3-you can use micro-array or PCR-array to find different expression pattern in patient and healthy person,also you can do deep seq or RNA-seq.
Best,
Mohsen
1. First of all, you should ascertain whether there are any miRNA diagnostic markers for the particular disease in your sample. Look up in the literature.
2. You also need to make sure that the samples used in the literature are the same as yours. You can have different markers for serum and tissue.
3. Depending on your sample, you might need to order a specific total RNA extraction kit with a protocol for small RNA extraction.
4. Find out other miRNAs to standardise your miRNA of interest. Remember to make sure that the normalising RNA is specific to your disease and sample/cell line. You may either look up literature, use public forums or run your own tests to find a normalising miRNA.
5. Once you extract small RNA, prepare cDNA for both miRNA of interest and the normaliser. After this, run qPCR and analyse your results. That should get you through.
Cheers!
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