Starting with <100 ng of degraded total RNA I would need to do RNA amplification before preparing ScriptSeq RNA libraries.
1. rRNA depletion using RiboZero (starting with ~100 ng degraded totalRNA)
2. Amplification of rRNA depleted RNA (starting with the ~5-10 ng RNA from 1.)
3. Preparation of random primed RNA-Seq libraries using ScriptSeq.
We have successfully generated RNA libraries using RiboZero and ScriptSeq from ~500 ng degraded Total-RNA.
Suggestions for a RNA amplification kit for step 2. would be appreciated!
Cheers,
Jakob
1. rRNA depletion using RiboZero (starting with ~100 ng degraded totalRNA)
2. Amplification of rRNA depleted RNA (starting with the ~5-10 ng RNA from 1.)
3. Preparation of random primed RNA-Seq libraries using ScriptSeq.
We have successfully generated RNA libraries using RiboZero and ScriptSeq from ~500 ng degraded Total-RNA.
Suggestions for a RNA amplification kit for step 2. would be appreciated!
Cheers,
Jakob
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