Hello everyone
I need an urgent help! :(
I have a year old samples of liver treated with regorafenib and other just the vehicle , after performing RT qPCR for them after one year (stored at -80c ) i got around 16 CT values for the vehicle groups and around 23CT for the regorafenib groups
can anyone help me to explain this, can drug actually affect the stability of RNA ????
To check the integrity of your RNA samples run your sample on agarose gel electrophoresis or bioanalyzer. If you see intact ribosomal RNA bands then your RNA is fine. This is just an indication because even if rRNA is partially degraded that doesn't mean that all mRNAs are affected.
Also make sure that your samples are treated with DNase to remove genomic DNA. If some of your samples have genomic DNA contamination then that will affect your qPCR results and will show lower Ct/Cq values compared to samples that don't have genomic DNA contamination
Best of luck
How was the RNA stored? in EtOH? In water? in Trizol? Were they processed and stored at the same time? also which gene were you analyzing? Not all housekeeping genes are good for all experimental setups
The best way to store you RNA is in water for no longer periods. However, can you explain more details please??
Can you also add more details, which method did you use to extract RNA? how much RNA you used for reverse transcription ...etc ?
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