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Old 11-30-2011, 11:20 PM   #1
nbogard
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Location: Singapore

Join Date: Apr 2011
Posts: 10
Default Custom SOLID Adapters for RNA-Seq

Has anyone here designed custom, SOLID-like adapters for preparing RNA-Seq libraries? Specifically, I'd like to use forward (F) and reverse (R), splint-like adapters (with poly-N overhangs) that include sequences used for the Illumina flow cell. Would this be as simple as just adding random-sequence overhangs to a sequence of my choosing?

As an example:

F...
5' - ACTGACTGACTG
3' - TGACTGACTGACNNNNN

R...
5' - PO4 - TTGGAACCTTGGAA
3' - NNNNAACCTTGGAACCTT

Also, do the SOLID adapters for RNA ligation consist of all DNA bases, or do they include some uracils (I've read posts with both)? And, besides the 5' PO4 on the R adapter, are there any other modifications required, or that aid, in the ligation?

Thanks
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