Sep 7, 2013
The changes to RNA-Seq library prep include:
1. The use of direct isolation of mRNA
The concern here is not knowing the integrity of the RNA isolated-but with cheaper sequencing and models to adjust for degradation it may not be a future issue. On the other hand this technique promises to reduce rRNA contamination.
2. Enzyme based fragmentation
3. Increased reliance on bead based purification.
I will say that the difference between gel slice purification and Ampure XP is the difference between success and failure of library prep in my case.