Storage of a plate for real time PCR - Ever stored your already prepared plate for real time PCR? (Oct/18/2007 )
I was just wondering. We have common real time machines and since one of them is not working, it's very very busy on the only other machine we have.
I know I can come back to work tonight at 10 and set up my whole plate (takes me at least 1 hr). But I'm really lazy and don't want to do that.
So I was wondering, I've heard somebody say that he freezes his plate when he needs to wait to use the real time machine. But I don't really trust the guy enough, so I want to double check if there are people here that have done that or have a very good way of dealing with something like this...
Other than coming in really late or really early!
Yep, freezing a plate for running it later (even a couple of days later) works just fine, I do it all the time!
When I'm running a couple of genes, I prepare all plates in the morning and store them in the fridge. It is important (I use SYBR green) that is is stored in a dark place, low temperature and off course not multiple freeze/thawing sessions of the SYBR green. Over night in the fridge works also fine, never tried a longer time than overnight however.
I also keep them (covered with alu-folie) ON in the fridge and it`s fine.
Thanks, I'm going to try it right away!
ensure that the qpcr master you are using contains a hot-start function. otherwise you may get problems with unspecific products (especially with sybr green).
i use the faststart universal probe master (rox) (Roche applied science) for my taqman assays or faststart universal sybr green master (rox) for sybrgreen assays on abi instrument (7500 fast). however i think it can also be used on other instruments as well (abi or others, check the pack insert). i stored my plates up to several days in the fridge and never had problems and no alterations compared to immediate runs after prep. Roche even say that you can store your plates at room temperature for up to 48hours when stored in dark.... so should be pretty stable.
my collegue who uses the abi sybr green FAST master said he had problems when storing the plate for longer times (he said 3 hours) - so i can't tell you something about stability of abi's master.
hope i could help you... please share your experiences...
Be sure the cover works well, otherwise the water inside wells can evaporate
Neyugn ,you are absolutely right.
Of course always apply enough pressure on the sealing foils and do not use the cheap ones they won't seal very well over prolonged times...