Go Back   SEQanswers > Applications Forums > Sample Prep / Library Generation

Similar Threads
Thread Thread Starter Forum Replies Last Post
RNA Integrity on Agilent's Bioanalyzer jvelotta RNA Sequencing 0 05-07-2013 01:15 PM
RNA integrity after SpeedVac RohanATX General 0 02-05-2013 01:20 PM

Thread Tools
Old 11-22-2017, 08:38 AM   #1
Junior Member
Location: Berlin

Join Date: Nov 2017
Posts: 2
Question RNA integrity number Drosophila

Hello @all
as we want to start an RNA-Seq analysis we try to determine the quality of our RNA (Drosophila heads) first. When we did this via Bioanalyzer an error occurred as expected. The measurement in a TapeStation results in a RIN though. I can`t explain why because still no 28S peak is visible. The RIN is the ration of 28S to 18S as I understood this.
Therefore I don`t know if I can rely on that. Does someone have experience with that?
Thank you!

ps: attached you see the trace of one sample with both machines
Attached Images
File Type: png Drosophila RNA_Bioanalyzer_9.png (68.9 KB, 25 views)
File Type: png DrosophilaRNA_TapeStation_9.png (92.5 KB, 21 views)
Elugop is offline   Reply With Quote
Old 11-22-2017, 10:50 AM   #2
Junior Member
Location: California

Join Date: Dec 2012
Posts: 4

It's a result of the 28S from insects being split. See this article:
ericchow is offline   Reply With Quote
Old 11-22-2017, 05:44 PM   #3
Senior Member
Location: US

Join Date: Dec 2010
Posts: 342

As consequence from what Eric wrote, one often has to eyeball the RNA integrity for insect samples on the Bioanalyzer (I am looking ratio of the rRNA peak size to the smaller debris fragments).
Your example trace looks great.
luc is offline   Reply With Quote
Old 11-27-2017, 04:41 AM   #4
Junior Member
Location: Berlin

Join Date: Nov 2017
Posts: 2

Thanks - I try it with these samples now. Fingers crossed
Elugop is offline   Reply With Quote
Old 11-27-2017, 08:02 AM   #5
Junior Member
Location: England

Join Date: Nov 2017
Posts: 1

I'd suggest using the Bioanalyzer over the Tapestation in the future. It gives a more granular electropherogram, whereas the Tapestation directly images the gel image from the run. This results in a less detailed electropherogram. Your trace looks good, but the RIN/RINe values are effectively redundant for insect material. Eyes are the best measure.

The link above is a great explanation!
HarbsSM is offline   Reply With Quote

Thread Tools

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off

All times are GMT -8. The time now is 03:08 PM.

Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2018, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO