Hi, I am transfecting mammalian cells with a miRNA mimic. After the 24-48hr transfection incubation I proceed to RNA isolation. My question is must the RT-PCR be run straight after RNA isolation or can the RNA be stored at -80 for some time before proceeding to qPCR and gene expression effects from the miRNA mimic will still be observed?
you absolutely can save your RNA at -80 and analyze later, snap freeze if you like.
Yes, RNA can be stored at -80 degrees C but several freeze and thaw can degrade the quality of RNA. It is preferred to aliquote RNA in small volumes and use if want to use it several times for further experiments.
June Leslie i would recommend you to prepare cDNA as soon as you isolate RNA and then freeze the remaining RNA in -80 because you freezing and thawing will decrease the efficiency of of RNA as cDNA are more stable than RNA.
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