Human blood cells stabilised by RNA-Later to perform DNA expression studies. It is a good way to store white blood cells to perform subsequently the isolation of RNA? Or is it preferable to store the pellets by a snap-freeze and without any solution?
I have used RNA-later in so many ways, always with good results. My recommendation would be spin the cells down to a small pellet, then aspirate any supernatant, then resuspend in a small amount (
Thanks Robert, but after the freeze storage at -20°C have you any recommendation to prepare the cells to the RNA extraction?
Sure, you can do a few different things. The easiest would be to spin the cells down again (gently, about 200xg) and aspirate the RNA Later off the pellet. Then get an RNeasy kit from Qiagen, they sell mini-spin columns for individual samples, or if you have a lot of samples, a 96 well format, RNeasy 96. The 96 well format requires a special centrifuge or a pump and a special manifold to hold it. The miini coulmns require only a microcentrifuge. These kits have a lysis buffer that will dissolve the preserved cells and you process from there to get purified RNA. I also recommend doing the DNase treatment if you are using the RNA for qPCR downstream. Check out the Qiagen website. Your other, less expensive method is to use Trizol, a phenol based extraction method, but Qiagen kits are much easier to use if you can afford to buy them.
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