Errors in RIN calculation due to signals in 5S region

Question

I have Run 2 Samples (pkst1 and pkst2) of Total RNA extracted from bacteria. It should have small RNAs along with mRNAs. The samples were quantified with QUbit4.0 and have 28ng/ul. The Total Nano-prokaryote kit was used for quantification. Result showed that there is "unexpected signals in 5S region". No "RIN" value was estimated though rRNA-Ratio was 0.4 and 0.5. 
 I have Ran the baseline correction and Ladder result was good. 
 Pls suggest how to resolve the results and get RIN values? 
 
 Thanks 
 Ram

Cendy.D · Accepted Answer

Hi singhRam19, 
 Thank you for your question. I am sorry to hear that you are observing this issue. Please kindly reference this Agilent Knowledge Portal Article that will guide you in attempting to release a RIN value: RIN Not Calculated or NA for Total RNA Samples in the Agilent Bioanalyzer 2100 Expert Software . 
 If there are still questions after reviewing steps from the Agilent Knowledge Portal Article, I suggest emailing us your raw data file that ends in .xad to electrophoresis@agilent.com . 
 Please let me know if I can be of further assistance.

Errors in RIN calculation due to signals in 5S region

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