Dear Mighty fellows,
I am new to RNA-Seq. Right now I am optimizing the RNA fragmentation conditions. I have tried incubating RNA with NEB Next fragmentation buffer for 3-5mins. All resulted in around 150nt fragments. I just tried 1min and 2min. In attached is a picture of that. My question is why there is always a 150nt peak in all conditions. Would anyone like to comment on that?
The other question is the fragment distribution after 2 min incubation too wide for RNA-Seq. I am making library for 100bp pair-end sequencing.
Your help is greatly appreciated.
I am new to RNA-Seq. Right now I am optimizing the RNA fragmentation conditions. I have tried incubating RNA with NEB Next fragmentation buffer for 3-5mins. All resulted in around 150nt fragments. I just tried 1min and 2min. In attached is a picture of that. My question is why there is always a 150nt peak in all conditions. Would anyone like to comment on that?
The other question is the fragment distribution after 2 min incubation too wide for RNA-Seq. I am making library for 100bp pair-end sequencing.
Your help is greatly appreciated.
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