[BioC] Rsubread for paired end SOLID sequencing

Wei Shi shi at wehi.EDU.AU
Sat Jul 27 04:20:23 CEST 2013


Dear Simon,

Yes, the F3/F5 protocol is supported by Subread. You will need to set PE_orientation="ff" when you run align or subjunc functions. This will tell the alignment programs that the two reads from the same pair are on the same strand and they can then be handled properly.

Sorry the documentation did not provide enough details about how to map such reads. I have just added more descriptions to this to the help pages and user guide to make them more clearer.

Best wishes,

Wei

On Jul 27, 2013, at 6:31 AM, Simon Coetzee wrote:

> I have some paired end SOLID sequencing data, the protocol using F3/F5
> fragments (not the simpler F3/R3).  Both fragments come from the same
> strand.  Some of the nuances are explained here:
> http://www.biostars.org/p/48597/
> 
> Having read the manual for Rsubread, its clear that it can handle SOLID
> sequencing, colorspace reads, and paired end sequencing.  However, is the
> F3/F5 protocol directly supported, or will there have to be preprocessing
> of the F5 fragments?
> 
> Simon Coetzee
> --
> Centro Regional de Hemoterapia
> Hospital das Clínicas da Faculdade de Medicina de *Ribeirão Preto
> *Universidade de São Paulo
> 
> -- 
> 
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