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Forum: RNA Sequencing 03-10-2011, 02:05 PM
Replies: 36
Views: 22,837
Posted By irit
I have used this kit a few times. For really...

I have used this kit a few times. For really effective removal of ribosomal RNA (as seen in bioanalyzer track) I had to do 2 rounds of ribodepletion. I had about 20% recovery (according to nanodrop,...
Forum: Sample Prep / Library Generation 03-09-2011, 09:27 AM
Replies: 268
Views: 98,801
Posted By irit
RNA-seq from single cells?

Does anyone here have experience with this? Does anyone know of a good protocol with consistent results?
Forum: RNA Sequencing 03-03-2011, 09:29 AM
Replies: 23
Views: 16,280
Posted By irit
If you are doing PolyA enrichment that would be...

If you are doing PolyA enrichment that would be the source of 3' bias. I don't know if there should be bias if not doing this (say, ribodepletion or dsn from total RNA).
Forum: RNA Sequencing 02-18-2011, 08:20 AM
Replies: 7
Views: 2,598
Posted By irit
A temperature drop should have the reverse effect...

A temperature drop should have the reverse effect - digestion of secondary structures.

Have you tested the enzyme to see if it is still active? Is it possible your incubation temperature is too...
Forum: RNA Sequencing 02-01-2011, 08:27 AM
Replies: 36
Views: 22,837
Posted By irit
Have you tried running your ribo-depleted RNA on...

Have you tried running your ribo-depleted RNA on Bioanalyzer? You won't be able to get a proper concentration reading but you should be able to see the size distribution of your RNA.
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