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Old 02-07-2013, 09:09 PM   #1
megalodon
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Location: Pennsylvania

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Default RNA quality - many peaks

Hi,

we recently extracted total RNA from mouse tissue (that had a bacterial infection). The bioanalyzer trace for the RNA sample is attached. Can anybody explain what all the peaks are in the trace? Could this be due to the fact that we probably co-extracted eukaryotic (mouse) and prokaryotic (from infection) simultaneously? If that is the case, is there anything that could be said about the quality of this sample.

Illumina recommends to use RNA with an RIN of at least 8. This sample has a RIN of 6.5 but I'm not sure if the RIN in this case accurately reflects the quality of the sample.

Thanks in advance for your help.

Daniela
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File Type: pdf RNA_trace.pdf (158.5 KB, 68 views)
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Old 02-25-2013, 12:56 PM   #2
MTTom
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I had an issue similar to this that I was able to fix it by diluting the sample. The bioanalyzer is highly sensitive to concentration and the Pico chip that you used is good between 50 and 5,000 pg/ul. Your concentration is estimated at 157,802pg/ul. I would dilute the sample way down and do not trust the results (RIN, concentration, etc...) if the reported concentration is outside of the 50-5,000pg/ul range.

On the other hand I was running Eukaryotic Total RNA and so it might be different...

Good Luck!

Tom
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