RNA-Seq and 5S
I am working on an RNA-Seq project and on epibio website under species compatibility it states for Mycobacterium tuberculosis "will not remove 5S". Looking at Agilent results I have to agree b/c we see a very distinct peak for the 5S region after using Ribo-Zero Magnetic Kit (Gram-Positive Bacteria) Cat. No. MRZGP126. 23S and 16S are gone. My concern is that a large portion of our reads from sequencing will be 5S region and we will reduce reads for the important stuff. Has anyone been successful at removing the 5S for such an experiment? Or do you just sequence more to achieve the desired depth?
I even see my library size range shifted. Usually I expect more reads >200bp, but in this case I have an increased distribution of fragments <200bp. I believe this to be the 5S region being fragmented even smaller and being carried through the rest of Library construction protocol. SoliD RNA-Seq standard protocol. I am very hesitant to proceed with sequencing. Does anyone have a similar experience?
Thanks.
Originally posted by Olaf Blue
View Post
I even see my library size range shifted. Usually I expect more reads >200bp, but in this case I have an increased distribution of fragments <200bp. I believe this to be the 5S region being fragmented even smaller and being carried through the rest of Library construction protocol. SoliD RNA-Seq standard protocol. I am very hesitant to proceed with sequencing. Does anyone have a similar experience?
Thanks.
Comment