Storage of RT-PCR products - (May/18/2005 )
I'm performing RT-PCR for the first time and was wondering what is the best way to store my RT products? Ideally I will be running PCR on my RT products right away but if PCR doesn't work I will need to use my RT products again. One postdoc in my lab said I can store my RT products at -20ºC (for how long?) and the other postdoc says 4ºC and to never freeze them. What do you guys normally do and for how long? Also, how long may I store my total RNA and at what temp? I usually store at -70ºC. Also, how many times may I freeze/ thaw these samples before they are severely degraded. Thanks
Either 4C or -20C is OK. For long-term storage, I store at -20C for several years without problem.
I work a lot with RT-qPCR. When we work with RNA, our enemy is RNase. Unfortunately, it exists every where, on your skin, in every cell, and it stays active for a long time!
1.To keep your RNA away from degradation by RNase guarantees the quality of RNA. So, keep your pre in -20C, but better in -80C for long storage.
2.Preserve RNA in RNase inhibitor soultion is also recommended. I use RNAsecure.
Before you perform RT reaction it is better to check if the RNA is still in good integrity.If the RNA is already degraded your cDNA will not be a good reflection of the RNA : (
If you are interested in more background info or any practice tips, site of Ambion, Qiagen or Agilent may provide you more information.
Good luck and have a lot of fun!