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I am trying to extract RNA from mice ears and for some reason I don't have RNA when I perform the electrophoresis. I directly cut the ears and I put it in a tube with a bead and trizol. then I place it on ice and I use a mash machine for tissue lysis for 8 minutes and 4 ºC. After that, I perform RNA isolation according with a protocol using chloroform, isopropanol and ethanol.

I think the problem could be the extraction. Do you have any suggestions?

Thanks.

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  • $\begingroup$ Do you see a pellet when you precipitate the RNA? $\endgroup$ – Chris Feb 10 '16 at 15:24
  • $\begingroup$ yes I saw a quite big pellet, but when I do the electrophoresis using a positive control, I suposse to see either two bands or a smears if it is degraded and it doesn't appear anything like that. $\endgroup$ – Bio Feb 10 '16 at 15:27
  • $\begingroup$ Let us know the composition, buffers, running conditions (voltage etc.) of your gel. $\endgroup$ – CKM Feb 10 '16 at 15:52
  • $\begingroup$ that's not the problem, since I use the samples to do a qPCR, I have used taqman, SYBRgreen, two different machines and I dont get amplification, but I do get in a previous sample used as positive control. So the problem is I don't have RNA in my samples. (the gel it was just to check if I have RNA and it also works with a positive control but not with my samples) $\endgroup$ – Bio Feb 10 '16 at 15:55
  • $\begingroup$ @Bio What were the A₂₆₀/A₂₈₀ and A₂₆₀/A₂₃₀ ratios? The problem could be the beads as well. Were the beads sterilized properly and kept in RNAse free conditions (in DEPC water)? $\endgroup$ – WYSIWYG Feb 11 '16 at 4:03

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