Has anyone had problems with low RNA recovery after chemical fragmentation for mRNA seq? A collaborator says that he loses ~90% of his material when he fragments chemically. He says that covaris shearing of RNA provides a much greater yield. Has anyone else had the same experience?
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Illumina kits uses chemical fragmentation for mRNA seq and they didn't see any problem with the yield of and so i don't think it is chemical fragmentation leads to low yield. Try to adjust the time of the fragmentation (typically 5 minutes). -
What kit was used for chemical fragmentation?
I used a commercial kit for fragmentation. The time was 15 mins. I tried fragmenting for 10 mins and increased it by 1 min up to 15 mins. I got the same result, all the RNA had been fragmented to below 100bp. I won't use this kit again.Comment
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The immune system’s power comes from its genetic diversity, allowing myriad threats to be neutralized through first recognizing foreign antigens. That diversity is also what makes the immune system so difficult to study. Recent advances in sequencing technology and computational biology, however, are giving researchers new tools to understand immune responses and immune-related diseases in greater detail.
This convergence of genetics, immunology, and computation...09-01-2026, 05:41 AM
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