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Old 10-24-2019, 12:59 AM   #1
nboom
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Location: London

Join Date: Oct 2019
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Default high rRNA after poly(A) selection

Hi,

About half our samples post library prep (poly(A) selection) + sequencing had around 50% rRNA content. The samples were re-prepped in the facility but the rRNA contents were similar a second time.

The facility now argue that the rRNA "contamination" is sample related and not due to failed library prep. Since the poly(A) selection enriches for all RNAs that have a poly(A) tail, mostly mRNAs, rRNA removal should have been effective? What can have gone wrong?

The tissue is skin and we used mirVana kit to extract RNA (perhaps not optimal - could the inclusion of small RNAs caused an issue in the library prep?). All our samples had RIN of 8 and above.

Thanks in advance!
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Old 10-25-2019, 02:56 AM   #2
Michael.Ante
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Default

Hi,

What library prep have you used?
Do you differentiate between MT-rRNA and nucleic rRNA? The mitochondrial rRNA molecules have a polyA tail and are therefore detectable after polyA selection.

Cheers,

Michael
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library preparation, poly a, rna sequencing, rrna contamination

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