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Old 09-17-2015, 03:51 PM   #6
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Location: Bioo Scientific, Austin, TX, USA

Join Date: Jun 2012
Posts: 119

Definitely use more starting RNA if you can. Worst-case scenario is you have more rRNA reads, but a library with high rRNA contamination is better than no library at all.

5 uL is also a pretty low input volume for RNA-Seq, I think typical kits are more like 15 uL. If your facility accepts completed libraries you may want to think about doing the preps on your own so you could have a little more flexibility with input amount and volume. This would also probably be cheaper, and would help you learn more about the library prep process. Also, the Ribo-Zero is probably one of the most expensive parts of your sequencing project, so it would be nice to be able to use the samples you have already prepared.
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