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Old 03-05-2014, 04:45 PM   #1
Seqwork
Junior Member
 
Location: NY,USA

Join Date: Mar 2014
Posts: 4
Question Low library yield

Hello Everyone,
I am new person to RNA Seq work.I made libraries from FFPE RNA as per protocol.RNA input was from 0.5-1ug for different samples.
But i can hardly see any yield on bioanalyzer and nanodrop readings are also 10ng/ul or below.
This is disappointing for me.I have been careful.I don't know where i lost my nucleic acid.One thing which i can remember about Ribosomal RNA removal beads step ,those beads were really difficult to dissolve.
Any suggestions will be welcomed.
Thanks
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Old 03-06-2014, 08:56 AM   #2
kwaraska
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Location: Boston,MA

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10ng/ul for an RNAseq library is not low-it is in fact quite good.

What does your bioanalyzer trace show?
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Old 03-06-2014, 11:57 AM   #3
Seqwork
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I made libraries twice.Both time nanodrop reading were 10ng/ul or below.
But i checked my libraries only first time on bioanalyzer, and i can see something at right size around 230-260 but very small peaks.
I have not checked my libraries second time at bioanalyzer, as i thought again i will not see anything.
someone suggested me that i should see at least 30ng/ul at nanodrop, but obviously bioanalyzer is more accurate.
what can i do to improve my yield.As i m planning to start it again.
Thank you for replying.
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