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Old 05-10-2011, 02:22 PM   #1
Location: san diego

Join Date: Nov 2009
Posts: 14
Default smallRNA seq input amount question

We have done many RNA seq experiments, but this will be our small RNA seq experiment. We are following small RNA library preparation using Total RNA sequencing kit. The protocol suggests either using total RNA or enriched small RNA for the library preparation. For the enrichment protocol we do not have sufficient total RNA amount (<2ug). So other option is to start the library prep with total RNA. Has anyone prepared small RNA library using total RNA as input. Can anyone suggest which is the best way to go - enrich small RNA with the available total RNA or use total RNA as input for library prep.

Please advice
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Old 05-12-2011, 09:17 AM   #2
Bioo Scientific
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Location: Austin, Tx

Join Date: Oct 2009
Posts: 99

Hi Sanush,

If you are using the NEXTflex™ Small RNA Sequencing Kit you can use 1 ug of total RNA or small enriched RNA from 1 - 10 ug of total RNA. Total RNA is fine for most cases. Enriching helps remove a lot of your ribosomal bands which will ensure more usable reads. How many lanes are you trying to run with your < 2ug sample?
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