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Old 08-20-2013, 01:21 PM   #1
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Location: Edmonton

Join Date: Aug 2013
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Default How important are A260/A230 and 28S/18s ratios are for RNA to be used miRNA deepseq

I did RNA isolation by TRIzol method from canola plants. When measured by NanoDrop 1000 the A260/A280 ratios for RNA are between 1.7-2.0 while the A260/A230 ratios are between 0.7-1.5. Though the RIN are quite high for all samples, ranging from 9.5-9.8, while the 28S/18s ratio is bit on the higher side, i.e. something between 2.0-4.8.

I am worried about the lower and higher A260/A230 and 28S/18s ratios! I saw a forum where people said that plant RNA tend to show higher 28s/18s ratio. Is it because of nuclear 25s RNA also coming in. Just a wild guess from my side.

Also, the concentration by Bioanalyzer is more than 90 ng/microlitre for most of samples.

Finally, is it safe to use this RNA for cDNA synthesis or should I do the isolation again??
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