[Bioc-sig-seq] trimLRPatterns - adaptor trimming
Martin Morgan
mtmorgan at fhcrc.org
Wed Aug 18 00:25:38 CEST 2010
On 08/17/2010 02:55 PM, Steve Lianoglou wrote:
> Hi Jess,
>
> On Tue, Aug 17, 2010 at 5:15 PM, JASREET HUNDAL <jasreeth at gmail.com> wrote:
>> Hi Martin & Harris
>>
>> It looks like the bioconductor packages that I am using are updated:
> ...
>> Also, this is error I get while exporting to fasta file:
>>> fa <-sread(trimmed)
>> Error in slot(object, "sread") :
>> no slot of name "sread" for this object of class "DNAStringSet"
My bad. The script Jess mention says
trimmed <- trimLRPatterns(pcrPrimer, subject=sread(seq))
and then trimmed is a DNAStringSet (try class(trimmed)), and writeFastq
doesn't make sense. With
trimmed <- trimLRPatterns(pcrPrimer, subject=seq)
it's a ShortReadQ (i.e., including quality) and writeFastq(trimmed,
"/path/to/file") should work. Explanations Steve offers below still
apply to th DNAStringSet.
Martin
>
> The "sread" method is used to get the *reads* (which would be a
> DNAStringSet object ) out of a ShortRead* object. The ShortRead* class
> stores the reads, as well as other information about the reads (phred
> scores, etc.)
>
> It looks like your "trimmed" object is already a DNAStringSet -- so
> calling sread() on a DNAStringSet isn't really meaningful ... you
> would just get itself.
>
> In short, the "trimmed" object you have appears to the the
> DNAStringSet you are looking to get by using the "sread" function.
>
>> And while exporting using writeFastq:
>>
>>> writeFastq(trimmed,"trimmedseqs")
>> Error in function (classes, fdef, mtable) :
>> unable to find an inherited method for function "writeFastq", for
>> signature "DNAStringSet"
>>
>> It would be great if you could suggest some other method to export the
>> trimmed object.
>
> Here you have the opposite problem.
>
> You "just" have a DNAStringSet -- no other metadata about your reads
> that a ShortRead* class holds, like phred scores, etc.
>
> The writeFastq file expects to write your object as a FASTQ file.
>
> If you have a DNAStringSet (which is what your "trimmed" object is),
> you can only really hope to write a FASTA file, in which case you can
> use the "writeFASTA" function (or the write.XStringSet function that
> Martin suggested) from the Biostrings package (which is already
> loaded).
>
> Does that make sense?
>
> So again (in summary (?)):
> the problem you are having is that you think that your "trimmed"
> object is something it's not.
>
> I'm not sure how you got it, or where it came from, but it is "just" a
> DNAStringSEt -- which is simply a number of sequences (reads).
>
> You can't write it as a fastq file (with writeFastq), and there's no
> need to get the "reads" out of it (with the sread function) because
> that's what they already are.
>
> Hope that helps,
>
--
Martin Morgan
Computational Biology / Fred Hutchinson Cancer Research Center
1100 Fairview Ave. N.
PO Box 19024 Seattle, WA 98109
Location: Arnold Building M1 B861
Phone: (206) 667-2793
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