Protocol Online logo
Top : Forum Archives: : siRNA, microRNA and RNAi

cDNA stability after miRNA stem-loop RT - (Mar/08/2008 )

I'm currently using ABI's stem-loop RT system for real-time analysis of microRNA. In their protocol, cDNA is diluted 1:10 before going into the real-time reaction but they say you can store it, undiluted, at -20C overnight and that longer periods of storage have not been tested. This struck me as odd...why would they say this if it is generally understood that cDNA is quite stable at -20C, even for years. Is there some reason that it would be different for this miRNA assay? Also, why do they say to store it undiluted? I was really hoping to be able to save my diluted cDNA (what I don't use for my current miRNA assays) for possible future sounds like they're saying this can't be done? I'm going to give it a try myself to see if I get different results after week or a month of storage at -20C, diluted, but does anyone have any ideas about this? Thanks! smile.gif



I remember a paper related to this, but I still not sure for its results
I have tested my polyA tailing based cDNA for miRNA, seem stable after year of storage..


Thanks rye. That paper has disturbing implications for my work but well....better to know now, right? :-)