I came across few genes which are fulfilling the characteristics of both miRNAs and lncRNAs through prediction as well as validation tools online. I would like to know some insilico as well as experimental method for validating these genes in-vitro. Can anyone suggest me methods other than luciferase assay and mimics method!
I personally use qPCR to validate any novel transcript.
I hope that help
Hi,
All in silico studies require experimental validations, therefore you cannot validate them bioinformatically.
qPCR is the best way to verify them initially. But please take note that you have to overexpress the pre-miRNA sequences in the cells and let Dicer process them. If an increase in mature miRNA level is observed, you can confidently claim that a new miRNA has been detected and validated.
For predicting the pre-miRNA sequence you can use the databases listed below:
miRNAFOLD, miPred, mFOLD, CID-miRNA, FomiR.
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