Hi all,
I extracted RNA from Drosophila heads, and want to do RNAseq on them. I used Agilent's Bioanalyzer to assess RNA quality, and found an expected double peak at the 18S region; I read that this is common in insects, and due to 28S denaturation, so I am not worried about that. However, I think that due to this double-peak all my RINs are N/A, and I want to ask Drosophilists or insect scientists with experience how they assess RNA quality for library preparation.
I attached an example sample to this thread; all my samples are similar, even though the RNA concentration varies greatly, and the attached sample is my highest concentration (~400 ng/ul); my lowest concentration was about 30 ng/ul, but, again, shows the same characteristic double-peak and no 'shoulder' in the fast region.
Best,
Pat
I extracted RNA from Drosophila heads, and want to do RNAseq on them. I used Agilent's Bioanalyzer to assess RNA quality, and found an expected double peak at the 18S region; I read that this is common in insects, and due to 28S denaturation, so I am not worried about that. However, I think that due to this double-peak all my RINs are N/A, and I want to ask Drosophilists or insect scientists with experience how they assess RNA quality for library preparation.
I attached an example sample to this thread; all my samples are similar, even though the RNA concentration varies greatly, and the attached sample is my highest concentration (~400 ng/ul); my lowest concentration was about 30 ng/ul, but, again, shows the same characteristic double-peak and no 'shoulder' in the fast region.
Best,
Pat
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