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Old 11-21-2012, 03:26 AM   #1
Helleaks
Junior Member
 
Location: Norway

Join Date: Aug 2012
Posts: 1
Default Scriptminer Small RNA-seq library troubles

Hi!

I have a question regarding my small-RNA library results on my bioanalyzer.
Recording to the scriptminer protocol you should get a peak around 150 bp (including adapters and miRNA (18-40 nt).
Attached you find my bioanalyzer results on CD4 cells (RIN 9.0)
Library 1 is totRNA from manufacturer. THis looks just like it should be. The 3 next libraries is my CD4 human cells. Here i have DNA around 150 bp, but i also got a peak around 200 bp. Is this normal? Is it just larger rna fragments in these type of cells?
I also did a small-RNA library prep in wheat cells. Here my results showed a peak that had an area from 150 bp- 300 bp) Should i just go further with the rest of my samples?
Attached Files
File Type: pdf SKMBT_C36012112113090.pdf (229.4 KB, 30 views)
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Old 11-27-2012, 09:00 AM   #2
cascoamarillo
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Location: MA

Join Date: Oct 2010
Posts: 158
Default

Hi,
Just a couple of questions; how do you get the Total/miRNA? I'm not sure about Scritpminer initial steps protocol, do you size select or use another kit (mirVana??).
The only way to know (I think) what you have in there is by colony screening (before sending to the Illumina machine); so you'll have an idea of potential contaminats in your library (rRNA...) and its proportion.
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