Questions tagged [deseq2]
Deseq2 is an R package for analyzing RNA sequencing data
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DESeq2 throwing error while normalizing raw microarray expression data due to presence of negative values
This question was also asked on Biostars
I am trying to download and analyze a miRNA expression dataset from NCBI GEO (GSE25631). I specifically want non-normalized ...
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RNAseq alignment: best practices for aligning to multiple isoforms?
I have Illumina RNAseq data and would like to maximize my power to find candidate genes that are differentially expressed genes between experimental conditions.
Many of my (de novo assembled and ...
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Differential gene expression: how do I correctly compare three groups to each other?
I'm trying to figure out the right way to do differential gene expression analysis with a discrete variable with three groups. The context is, I want to assess differential expression as a function of ...
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Batch correction in differential expression analysis
I have sent two sets (two batches in matter of sending for sequencing) of different samples (plasma) to small RNA-seq to Qiagen company
This is how my meta data look
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Manually calculating log2 fold change values from DESeq2 normalized counts
This question was also asked on Biostars and Bioconductor Support
My aim is to perform gene clustering based on RNA seq datasets (raw expression values) from NCBI database. After clustering I realized ...
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DESeq2 for large number of samples takes too much RAM
I am trying to run a very large number of transposase-accessible chromatin (ATAC)-seq samples through DESeq2 to find peaks of differential chromatin accessible across my study genome.
I have about 100 ...
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DESeq2 and EdgeR
I am new to using both DESeq2 and EdgeR in Bioconductor used for transforming my RNA expression data.
However, I am struggling to understand their specific purpose, differences between them and ...
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Filtering reads greater than 5 from HT-seq count files
I have some raw counts from HTSeq after aligning with hg38 human reference genome. I want to do filtering in a way that the filtered count files should have the same number of lines. The reason behind ...
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Creating multiple phenotype datasets using bootstrap method "Bootstrap-samples-by-column-of-a-data-frame-in-r" for DEG analysis
I am working on a datasets and after some discussion with my group, we doubt that maybe one or more of our controls are different than the other controls. The motivation is to see if one or more ...
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nested samples in design DEseq2
I'm running a DEG analysis with Deseq2 by specifing the following design that includes 12 samples from 6 pts in 4 different conditions:
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Basic RNA Differential Expression in R
I have two matrices, one for individuals before treatment and one for the same individuals after treatment. Both matrices are raw read counts of RNA expression.
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deseq2 single factor design output interpretation
This is regarding the single factor design For example if I have Age or other continuous numerical variable how to provide that into the design formula.
For this post do i need to 'You could ...
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deseq2 makeContrasts function
This article talked about various deseq2 designs etc. One of the designs I would like to use is explained as this:
Control versus treatment average
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deseq2 full and reduced model interpretation
I would like to know how to interpret the output of the formula although i been thorough loads of literature but I'm not confident yet.
So this is my full model I'm running
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Different results of DESeq2 in different ways of comparison
I have different treatments (infected and control ones). When using DESeq2, I have different results in two ways of comparison.
first way:
factor: treatment, factor level1: Moribund Infected, factor ...
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What is the difference between Normalized Expression in EdgeR vs DESeq2?
I am trying to access the normalized expression in both edgeR and DESeq2, yet the results are different. Does anyone know why?
How to get normalized expression using edgeR:
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baseMean threshold
I have an RNA-seq dataset and I am using DEseq2 to find differentially expressed genes between the two groups. I used pre-filtering to remove any genes that have ...
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Error in DESeq2 analysis with dimnames
I'm getting the following error in a DESeq2 analysis and I can't seem to figure out the issue:
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LRT test in deseq2 on multiple condition
I have some doubts when i'm running LRT
This is my condition data-frame small subset
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How to input data and metadata from NCBI for RNA-Seq analysis in R
I am quite new to R programming so I hope my question is a clear one and not too confusing.
So, I have downloaded some .txt and .csv files containing the raw count data (I am supposed to find out ...
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How does DESeq2 "collapseReplicates()" function work on read counts data?
Comparing read counts from an RNA-seq experiment for two select genes before and after using DESeq2's collapseReplicates() and ...
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Batch Effects that are Close to Collinear with Main Effect
Two years ago, I started working on a project uses both RNAseq and ATACseq. It's supposed to be a simple Healthy Control (HC) vs disease study. The sample collection was done in 2 phases, and it was ...
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How to remedy a DESeq2 collapsing technical replicates error?
Goal: To ensure "the sum of the counts for [my samples] is the same as the counts in the [samples] columns in ddsColl" after collapsing replicates using ...
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Use of "baseMean" in DeSEQ2
The basemean is described as the "mean of normalized counts of all samples". My question is, how would we interpret differences in basemean between genes, pertaining to reliability and such? ...
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Visualizing top expressed genes
I have a spreadsheet of CPM values
I want to visualize the top 20 genes expressed across my samples
This is how my data look like
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Best R program to use for RNA seq analysis
We have an RNA seq taken from a the same tissue in two different developmental stages. We have one gene, with the number of reads from stage A and stage B, and we want to see which genes are ...
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The correct design for time series experiment
I have a treatment and control in two time points like this
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How to simulate replicates for DGE analysis?
I am prototyping with data visualization of DGE results, and I would like to work on the analysis pipeline before the real data is available. Currently, I only have 3 samples for wild type and 1 ...
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Help with performing PCA analysis on data from 3 different datasets of normalized counts data for RNA-seq experiment
So, I have limited knowledge of R but I need to do a PCA analysis of 3 different datasets of gene expression as a result of combined growth or mono-culture growth.
The 3 different datasets I performed ...
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Understanding the different designs in DESeq2
I am using DEseq2 and trying to understand the results obtained using different models.
I have a data design with 2 genotypes and 2 time points.
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DEseq2 contrast in two time points
I need help with some clarifications please
I have control versus treatment in two time points like
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Differential miRNA expression using RPM
I have microRNA (miRNA) expression data in RPM. I would like to do differential gene expression between two groups.
How can I do this?
I have considered edgeR and DESeq2 in R, but it looks like they ...
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Adjusted P-value from DESeq2
What does the padj really mean? I believe it's a BH method, where it attempts to quantify the likelihood of a "false positive" but I am not sure. Also, what does it mean if the resulting ...
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DESEQ2 Question about results()
I am currently learning to perform Differential Analysis via DESEQ2 R Package, and I believe I've made progress, able to format the data correctly [maybe] for DDS(). When I run the results function to ...
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Beginner with DESeq2 having issue with analysis
I am trying to run DESeq2 analysis on my featureCounts output counts table.
I ran the following code:
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DESeq2. Which is better: using the altHypothesis argument in the results function or manually filtering for your desired P-value and Log2FoldChange?
In DESeq2 you can identify your differentially expressed (DE) genes using the results function. I noticed people in my lab using the results() function with the minimum number of arguments supplied (...
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How does FDRtool work?
I have a question about using FDRtool. In the below code (on RNA seq data whose p values were acquired using Deseq2), the FDRtool was first used and thereafter p.adjust using the benjamini hochberg ...
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Calculating most abundant transcript from RNA-Seq data
vcf2maf uses VEP to annotate variants, and I believe selects the default Ensembl transcript to use for annotation. Sometimes the ...
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DESeq2 multiple treatments, multiple time points, multiple cell lines
Note: this question has also been asked on Bioconductor Support
I know there are a lot of questions asking similar things here on this forum and I checked the vingette and did a lot of other research ...
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Un-normalize DESeq2 counts
I obtained a matrix of RNA-seq count data that has been normalized by DESeq2's median of ratio method.
I know that DESeq2 wants to take in un-normalized counts, but I do not have access to those data.
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Bulk RNA-Seq Read Length Normalization across different samples
I have 20 samples out of which 14 are 100 bp in length and 6 are 150 bp. Is there a way to normalize the read length for cross-sample differential expression comparison? What would be the best way to ...
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Differential expression analysis for a subset of transcripts
I want to check the differential expression of a specific class of transcripts (say, long non-coding RNAs) using DESeq2. Now, I know that the normalization step takes into account the total number of ...
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Adding matrix as a confounder in EnrichmentBrowser
Brain data, particualrly human brain data have very heterogeneus cell types, so it's important to normalise by cell type/add proportoins of different cell types to the formula when performing ...
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the model matrix is not full rank : This is a classic question which a biologist face without clear understanding of the model design
Saw this answer biostar. Tried to make my metadata as such but still the
"Error in checkFullRank(modelMatrix) : "
This is my coldata
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What is a sensible number of gene/observations to explain PCA variance?
I am working with a set of RNASeq dataset. I have about 4000 observations (genes) on 20 samples and plotting a PCA I found the clustering doesn't vary much when I use different number of genes, but ...
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Question of Padj value (very high, infinite) got from dds
After I got dds from my count matrix by DESeq(), I use results() method to get Padj and log2 fold changes for volcano plotting,
then I found some gene's Padj values are infinite and the P-value is ...
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Why use "robust" FPKMs?
Both DESeq2 and edgeR have an FPKM/RPKM function that by default uses normalized library sizes ("robust" option in DESeq2). FPKMs have their own issues, but I thought the main benefit was to ...
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Is there an established method for comparing expression of groups of genes (gene sets)?
I have a figure in a paper where I show the log2(fold-change) (obtained with DESeq2) between two groups (based on genotype) for genes within a specific hallmark gene set, and a reviewer is asking ...
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