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-   -   How the sequential adaptation process works? (http://seqanswers.com/forums/showthread.php?t=77134)

wangwde 07-19-2017 02:03 AM

How the sequential adaptation process works?
 
Hi, saw some library kits using sequential adatation process, first to add adapter to the 5 ' prime and then 3'prime, just wondering how it works, many thanks in advance!

huguesparri 07-19-2017 10:34 PM

Hello,

This kind of protocol is typical of smallRNAseq.
It uses a truncated T4 RNA ligase 2 and a modified 3' adapter.
Trunc. T4 RNA ligase is unable to adenylate the 5' end of its substrate so it uses the modification of the 3' primer (rApp) to perform this step.
The second ligation uses a classical RNA ligase.

I wasn't aware of such protocols used for RNAseq...

wangwde 07-19-2017 10:43 PM

Thanks!
Swift Bio for example using this strategy to build their Libraries, for DNAseq and RNAseq. It should be different for DNA and RNA application, right?


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