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No 5S RNA on a QC gel

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#1 PhD28



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Posted 12 February 2013 - 12:11 AM


I recently did an RNA extraction it two halves. I compare the samples in each half directly. I need a lot of RNA, so I split each sample into three tubes after trizol harvest and then process them.

When I ran the gel I noticed that I am missing the 5S band in half of the samples, but the higher rRNA bands are fine and the microRNAs that I am examining also seem to be fine.

Has anyone got any suggestions?

I am heating my RNA to see if it is a problem with resuspension, but it seems unlikely that this would affect smaller RNA, as I would have thought that this would go into solution quicker. I use 30ul to resuspend each sample, moving it between each of the 3 tubes, so perhaps some RNA was left behind.

Any suggestiong would be appreciated because I can't think of an explanation for this!

Also the RNA doesn't look degraded, it is just that the lower rRNA band is missing in these samples.

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