Hi Farideh

You can use cel-miR-39 or whatever other spike-in control to check either the extraction of your RNA (by adding the spike in before) or the efficiency of the cDNA synthesis, by adding the spike-in after RNA extraction. I would recommend to use two different synthetic spike-in molecules if you wish to check for both quantification steps.

Best luck.

Paco

Hi Farideh

You can use cel-miR-39 or whatever other spike-in control to check either the extraction of your RNA (by adding the spike in before) or the efficiency of the cDNA synthesis, by adding the spike-in after RNA extraction. I would recommend to use two different synthetic spike-in molecules if you wish to check for both quantification steps.

Best luck.

Paco

 Hello Professor Francisco J. Enguita

Thank you for your answer and your recommend.

Hi Farideh

I think it is also possible to use cel-miR-39 in normalization, in order to normalize for some of the technical variability. We used it during normalization, together with endogenous controls. 

BR, Julia

hi

cel-miR-39-3p could be an ideal spike in miRNA control for even serum samples having relatively high PCR efficiency but it has to be added only after incubation with lysis reagent. I am using RNAisoplus as a lysis reagent along with miRNA carrier molecule, specially for serum in which there is less abundance of miRNA and chances of degradation is more due to RNase in serum volume used as 150-200ul. This spike in can be used along with negative control for miRNA extraction using RNase free water to check extraction efficiency as well as normalization of test sample apart from endogenous control for miRNA. Other option could be cel-miR-54 as a spike in miRNA.

all the best