RRBS question
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@alex-gutteridge-2935
Last seen 9.6 years ago
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On 21.01.2014 20:48, Gilgi Friedlander wrote: > Hi Alex, > > Sorry to bother you, but I have a question regarding RRBS analysis. > > I saw your post from last year, and also was wondering if bsmooth can > work well for RRBS data, as going to work on such data. > > I wanted to reply to the post on the mailing list, but didn't see > such an option. > > If you already have results, and can share your experience if one can > use bsmooth for RRBS, it will be great. And if not, if you have > recommendations for other tools. > > Thanks a lot, > > Gilgi cc'ing the list for future reference and in case anyone else has additional insights on RRBS analysis with Bioconductor: Hi Gilgi, I had mixed results with bsmooth and rrbs data when I tried it. Certainly the various smoothing parameters required tweaking before it would run successfully. I can let you know what worked for me, but I'd be lying if I said I arrived at them through careful experimentation. One also has to be cautious I think about the overall approach of whether smoothing makes sense when the data from rrbs is by definition non-contiguous. That said, bsmooth did run and did detect differential methylated regions for us which looked correct on deeper inspection. What bsmooth (by design I guess) does not detect are the single CpGs that seem to change between experimental groups, but sit within larger regions that clearly do not change. It's still an open question (in my mind at least) whether such sites are likely to be biologically significant or not. -- Alex Gutteridge
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Tim Triche ★ 4.2k
@tim-triche-3561
Last seen 3.6 years ago
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BiSeq is designed to address some of these issues, although I'm not certain whether the "single CpG" issue is one of them. Obviously it would be a good idea to be 100% certain when looking at individual DMPs that the CpG is not a SNP with varying frequencies between the groups being compared :-) --t On Wed, Jan 22, 2014 at 2:15 AM, Alex Gutteridge <alexg@ruggedtextile.com>wrote: > On 21.01.2014 20:48, Gilgi Friedlander wrote: > >> Hi Alex, >> >> Sorry to bother you, but I have a question regarding RRBS analysis. >> >> I saw your post from last year, and also was wondering if bsmooth can >> work well for RRBS data, as going to work on such data. >> >> I wanted to reply to the post on the mailing list, but didn't see >> such an option. >> >> If you already have results, and can share your experience if one can >> use bsmooth for RRBS, it will be great. And if not, if you have >> recommendations for other tools. >> >> Thanks a lot, >> >> Gilgi >> > > cc'ing the list for future reference and in case anyone else has > additional insights on RRBS analysis with Bioconductor: > > Hi Gilgi, > > I had mixed results with bsmooth and rrbs data when I tried it. Certainly > the various smoothing parameters required tweaking before it would run > successfully. I can let you know what worked for me, but I'd be lying if I > said I arrived at them through careful experimentation. One also has to be > cautious I think about the overall approach of whether smoothing makes > sense when the data from rrbs is by definition non-contiguous. That said, > bsmooth did run and did detect differential methylated regions for us which > looked correct on deeper inspection. What bsmooth (by design I guess) does > not detect are the single CpGs that seem to change between experimental > groups, but sit within larger regions that clearly do not change. It's > still an open question (in my mind at least) whether such sites are likely > to be biologically significant or not. > > -- > Alex Gutteridge > > _______________________________________________ > Bioconductor mailing list > Bioconductor@r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/gmane. > science.biology.informatics.conductor > [[alternative HTML version deleted]]
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@kasper-daniel-hansen-2979
Last seen 9 months ago
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Because RRBS data is non-contiguous, you basically have to use the 'maxGap' argument to do the smoothing on each of the contiguous groups. You probably also have to use 'local.correct=FALSE' as the algorithm is currently written. I have not yet experience with applying the algorithm to RRBS data myself. Best, Kasper On Wed, Jan 22, 2014 at 5:15 AM, Alex Gutteridge <alexg@ruggedtextile.com>wrote: > On 21.01.2014 20:48, Gilgi Friedlander wrote: > >> Hi Alex, >> >> Sorry to bother you, but I have a question regarding RRBS analysis. >> >> I saw your post from last year, and also was wondering if bsmooth can >> work well for RRBS data, as going to work on such data. >> >> I wanted to reply to the post on the mailing list, but didn't see >> such an option. >> >> If you already have results, and can share your experience if one can >> use bsmooth for RRBS, it will be great. And if not, if you have >> recommendations for other tools. >> >> Thanks a lot, >> >> Gilgi >> > > cc'ing the list for future reference and in case anyone else has > additional insights on RRBS analysis with Bioconductor: > > Hi Gilgi, > > I had mixed results with bsmooth and rrbs data when I tried it. Certainly > the various smoothing parameters required tweaking before it would run > successfully. I can let you know what worked for me, but I'd be lying if I > said I arrived at them through careful experimentation. One also has to be > cautious I think about the overall approach of whether smoothing makes > sense when the data from rrbs is by definition non-contiguous. That said, > bsmooth did run and did detect differential methylated regions for us which > looked correct on deeper inspection. What bsmooth (by design I guess) does > not detect are the single CpGs that seem to change between experimental > groups, but sit within larger regions that clearly do not change. It's > still an open question (in my mind at least) whether such sites are likely > to be biologically significant or not. > > -- > Alex Gutteridge > > _______________________________________________ > Bioconductor mailing list > Bioconductor@r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/gmane. > science.biology.informatics.conductor > [[alternative HTML version deleted]]
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Yes, this is exactly what I found. I ended up tweaking maxGap to 10kbp which seemed to give a reasonable result over most regions. Alex Gutteridge On 22.01.2014 16:29, Kasper Daniel Hansen wrote: > Because RRBS data is non-contiguous, you basically have to use the > 'maxGap' argument to do the smoothing on each of the contiguous > groups.? You probably also have to use 'local.correct=FALSE' as the > algorithm is currently written.? I have not yet experience with > applying the algorithm to RRBS data myself. > > Best, > Kasper > > On Wed, Jan 22, 2014 at 5:15 AM, Alex Gutteridge wrote: > >> On 21.01.2014 20:48, Gilgi Friedlander wrote: >> >>> Hi Alex, >>> >>> Sorry to bother you, but I have a question regarding RRBS >>> analysis. >>> >>> I saw your post from last year, and also was wondering if >>> bsmooth can >>> work well for RRBS data, as going to work on such data. >>> >>> I wanted to reply to the post on the mailing list, but didn't >>> see >>> such an option. >>> >>> If you already have results, and can share your experience if >>> one can >>> use bsmooth for RRBS, it will be great. And if not, if you have >>> recommendations for other tools. >>> >>> Thanks a lot, >>> >>> Gilgi >> >> cc'ing the list for future reference and in case anyone else has >> additional insights on RRBS analysis with Bioconductor: >> >> Hi Gilgi, >> >> I had mixed results with bsmooth and rrbs data when I tried it. >> Certainly the various smoothing parameters required tweaking before >> it would run successfully. I can let you know what worked for me, >> but I'd be lying if I said I arrived at them through careful >> experimentation. One also has to be cautious I think about the >> overall approach of whether smoothing makes sense when the data > from >> rrbs is by definition non-contiguous. That said, bsmooth did run > and >> did detect differential methylated regions for us which looked >> correct on deeper inspection. What bsmooth (by design I guess) does >> not detect are the single CpGs that seem to change between >> experimental groups, but sit within larger regions that clearly do >> not change. It's still an open question (in my mind at least) >> whether such sites are likely to be biologically significant or > not. >> >> -- >> Alex Gutteridge >> >> _______________________________________________ >> Bioconductor mailing list >> Bioconductor at r-project.org [1] >> https://stat.ethz.ch/mailman/listinfo/bioconductor [2] >> Search the archives: >> http://news.gmane.org/gmane.science.biology.informatics.conductor >> [3] > > > > Links: > ------ > [1] mailto:Bioconductor at r-project.org > [2] https://stat.ethz.ch/mailman/listinfo/bioconductor > [3] http://news.gmane.org/gmane.science.biology.informatics.conductor > [4] mailto:alexg at ruggedtextile.com -- Alex Gutteridge
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Thank you so much for all the help and important information! -----Original Message----- From: Alex Gutteridge [mailto:alexg@ruggedtextile.com] Sent: Wednesday, January 22, 2014 7:06 PM To: Kasper Daniel Hansen Cc: Gilgi Friedlander; bioconductor at r-project.org Subject: Re: [BioC] RRBS question Yes, this is exactly what I found. I ended up tweaking maxGap to 10kbp which seemed to give a reasonable result over most regions. Alex Gutteridge On 22.01.2014 16:29, Kasper Daniel Hansen wrote: > Because RRBS data is non-contiguous, you basically have to use the > 'maxGap' argument to do the smoothing on each of the contiguous > groups.? You probably also have to use 'local.correct=FALSE' as the > algorithm is currently written.? I have not yet experience with > applying the algorithm to RRBS data myself. > > Best, > Kasper > > On Wed, Jan 22, 2014 at 5:15 AM, Alex Gutteridge wrote: > >> On 21.01.2014 20:48, Gilgi Friedlander wrote: >> >>> Hi Alex, >>> >>> Sorry to bother you, but I have a question regarding RRBS analysis. >>> >>> I saw your post from last year, and also was wondering if bsmooth >>> can work well for RRBS data, as going to work on such data. >>> >>> I wanted to reply to the post on the mailing list, but didn't see >>> such an option. >>> >>> If you already have results, and can share your experience if one >>> can use bsmooth for RRBS, it will be great. And if not, if you have >>> recommendations for other tools. >>> >>> Thanks a lot, >>> >>> Gilgi >> >> cc'ing the list for future reference and in case anyone else has >> additional insights on RRBS analysis with Bioconductor: >> >> Hi Gilgi, >> >> I had mixed results with bsmooth and rrbs data when I tried it. >> Certainly the various smoothing parameters required tweaking before >> it would run successfully. I can let you know what worked for me, but >> I'd be lying if I said I arrived at them through careful >> experimentation. One also has to be cautious I think about the >> overall approach of whether smoothing makes sense when the data > from >> rrbs is by definition non-contiguous. That said, bsmooth did run > and >> did detect differential methylated regions for us which looked >> correct on deeper inspection. What bsmooth (by design I guess) does >> not detect are the single CpGs that seem to change between >> experimental groups, but sit within larger regions that clearly do >> not change. It's still an open question (in my mind at least) whether >> such sites are likely to be biologically significant or > not. >> >> -- >> Alex Gutteridge >> >> _______________________________________________ >> Bioconductor mailing list >> Bioconductor at r-project.org [1] >> https://stat.ethz.ch/mailman/listinfo/bioconductor [2] Search the >> archives: >> http://news.gmane.org/gmane.science.biology.informatics.conductor >> [3] > > > > Links: > ------ > [1] mailto:Bioconductor at r-project.org > [2] https://stat.ethz.ch/mailman/listinfo/bioconductor > [3] http://news.gmane.org/gmane.science.biology.informatics.conductor > [4] mailto:alexg at ruggedtextile.com -- Alex Gutteridge
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