I usually use RNAlater to isolate mRNA from human skin biopsies and make RT-PCR and have never had problems.
But this last time when I went to process the skin (tube +skin + RNA later at 4ºC) I saw the skin had changed color from transparent to blue. Could this affect to the mRNA?
Somebody knows why ??
Tx
Idk why but tissue stored in RNALater at 4*C is fine for at least a month,
http://www.sigmaaldrich.com/content/dam/sigma-aldrich/life-science/molecular-biology/migrationmolbio1/RNAlater-figure-2a.gif
Have you tried extracting the RNA and check its integrity?
Yes, I checked this link and that's what I knew, but as the tissue changed the color from transparent to blue, purple...
I have to said that work with RNAlater is wonderful.
I measured the concentration (not the integrity) of the RNA and I did a RT-PCR and it worked good.
Although why it changed the colour, that is what I do not know. Maybe the skin proteins reacted with the RNAlater... I do not know.
Anyway thanks a lot for your answer.
Yes RNALater is great
I feel that I should tell you if your RNA is degraded you can still amplify for qPCR. However certain conditions can be insensitive to RNA integrity
"In general we can point out, that amplification of long product over 400 bp is
strongly dependent on a good RNA quality, which should show at least a RIN of
5. Shorter qRT-PCR products, mostly used with the length of 70–250 bp, are more
or less ‘‘independent’’ of the RNA quality."
http://gene-quantification.org/fleige-pfaffl-rin-2006.pdf
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