Category: DiffBind

My ATAC-seq data returns a peak of 1bp for all the peak intervals when using DiffBind’s dba.count(summit=0). why is this happening?

My ATAC-seq data returns a peak of 1bp for all the peak intervals when using DiffBind’s dba.count(summit=0). why is this happening? 0 The binding matrix’s peak intervals (all of them) have a width =1 bp. I have set summit=False, summit=0, summit=True. All these parameters return 1 bp width. In fact,…

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why are all DiffBind tutorial’s ATAC-seq peak intervals 400 bp for all intervals?

why are all DiffBind tutorial’s ATAC-seq peak intervals 400 bp for all intervals? 1 I have followed DiffBind tutorial bioconductor.org/packages/release/bioc/vignettes/DiffBind/inst/doc/DiffBind.pdf I have found out that the returned count matrix’s peak intervals are always 400bp. Literally, all the peak intervals in the count matrix are 400 bp. I am wondering why…

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How to import a BED file to ChIPseeker in order to visualize genome coverage plot?

How to import a BED file to ChIPseeker in order to visualize genome coverage plot? 1 Could you please explain how to connect the ATAC-Seq peak results discovered by DiffBind to be imported to ChIPseeker to get CHIP peaks coverage plot? For example, I have the following bed file. chr14…

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How can I export raw count profile from DiffBind to be analyzed on DESeq2 separately?

How can I export raw count profile from DiffBind to be analyzed on DESeq2 separately? 0 I am performing some ATAC-seq analysis. I would like to export the output files as a raw abundance profile so that I can analyze them on DESeq2 separately. I am aware that DiffBind contains…

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DiffBind spike-in normalisation with varying amounts of spike-in chromatin

DiffBind spike-in normalisation with varying amounts of spike-in chromatin 0 Hi there! I am currently using DiffBind v3.2.7 to analyse some ChIP-seq data for RNA Polymerase III (RNAPIII). We have Drosophila spike-in chromatin in the samples that I would like to use in DiffBind to normalise the data. The problem…

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Help generating a correct linear model (DESeq2) for use in DiffBind3

Hello, I am trying to do a differential ATACseq analysis with DiffBind3, and need to build a design that DESeq2 will accept. I have tried several relevant models, but I have been unsuccessful in achieving the contrasts I desire, without DESeq2 throwing a “Model matrix not full rank” error. Any…

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New in DiffBind: dba.plotProfile() – Peak profiles and profile heatmaps

Hi, I am using the new plotProfile function of Diffbind to create some graphs, and get the following error with one subset of my data: dbObj$config$RunParallel <- TRUE profiles <- dba.plotProfile(dbObj) Generating report-based DBA object… Generating profiles… Error in value[3L] : profileplyr error: Error: BiocParallel errors element index: 2 first…

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dba.count reduces binding matrix, why?

dba.count reduces binding matrix, why? 0 db <- dba(samplesheet) This gives me: db 9 Samples, 138044 sites in matrix (193226 total) then I do db.cnt <- dba.count(db) db.cnt 9 Samples, 55893 sites in matrix I wonder why dba.count is removing more than half of the intervals? diffbind atacseq chipseq diffbind3…

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Disabling de novo DNA methylation in embryonic stem cells allows an illegitimate fate trajectory

The mammalian genome is characterized by widespread methylation of cytosine residues. After fertilization, however, both maternal and paternal genomes undergo extensive demethylation, reaching a low point in the blastocyst (1⇓⇓–4). The embryo genome is then remethylated by the activity of de novo DNA methylation enzymes (5). Mouse embryonic stem (ES)…

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Normalization and differential analysis in ATAC-seq data

Normalization and differential analysis in ATAC-seq data 2 Hello everyone! I would like to know if someone had experiences with normalization and differential expression on ATAC-seq data. After using MACS2 for the peak calling, how can we use Dseq2 or EdgeR on these datas? Someone try this? What is the…

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RPKM on TSS using DiffBind

RPKM on TSS using DiffBind 1 Hi everyone, I want to extrapolate RPKM value from Diffbind. Investigated regions are TSS (± 2.5kb) but around 3500 TSS are “lost” because merged. I have read that in the new version of DiffBind is possible to use dbObj_region$config$mergeOverlap with negative value to avoid…

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low number of significant peaks for one contrast

I am using Diffbind to call differential peaks on an ATAC seq dataset of four conditions (AW, BW, B, and C), and each condition has 2 replicates. One of my replicates (BW2) has low quality (low number of peaks detected by MACS2 compared to the other replicate, and low FRiP)….

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TMM normalization and sex effect

TMM normalization and sex effect 1 Hello to everyone. I have a doubt about TMM normalization. I’m comparing male versus female samples. Can TMM normalization be affected by the presence of more reads on chromosome x in the female group?? Thanks Francesca edgeR diffbind rnaseq tmm • 75 views It…

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Bioconductor – DESeq2

DOI: 10.18129/B9.bioc.DESeq2     This package is for version 3.10 of Bioconductor; for the stable, up-to-date release version, see DESeq2. Differential gene expression analysis based on the negative binomial distribution Bioconductor version: 3.10 Estimate variance-mean dependence in count data from high-throughput sequencing assays and test for differential expression based on…

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Bioconductor Forum

James W. MacDonald 57k 1 week, 5 days ago United States Answer: Biomart’s getBM returns no genes for an existing GO-term in grch38, and less the Michael Love 33k 1 week, 6 days ago United States Answer: Normalizing 5′ Nascent RNA-seq data to identify differentially expressed transcr Kevin Blighe 3.3k 2 weeks, 2 days ago Republic…

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DiffBind: dba.plotProfile() Change sample order

DiffBind: dba.plotProfile() Change sample order 0 Hi, Is it possible to change the order of samples when using dba.plotProfile()? I’ve tried defining them with the samples= parameter, but I haven’t been successful. I’ve made lovely plots (this is a great tool!), but instead of the default order (ex A, B,…

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