Categories
Tag: log2FoldChange
No genes mapped in clusterprofiler gseGO
Hello! I’m having issues generating an adequate geneList for running gseGO in clusterProfiler, using keytype = “GO” Similar issues have been described here: No gene mapped gseGO code is: gse <- gseGO(geneList = gene_List, ont = “ALL”, #ont one of “BP”, “MF”, “CC” or “ALL” OrgDb = OrgDb, minGSSize =…
Batch and Sample correction for downstream analysis using DESeq2
Hello everyone, I am an absolute beginner on sequencing analysis and DESeq2, so please forgive me for possibly mundane questions. I have tried to look up different methods, but couldn’t find a fitting answer yet. I am currently working with sequencing data derived from an Illumina sequencer. The data is…
Bioconductor GraphAndNetwork
Comment: flowCore >2.0 read.FCS results in $SPILLOVER error when reading certain FCS file by SamGG ▴ 320 Hi. rbio2021 has no spillover, so no problem with current flowCore. As you identified it, the spillover matrix of rbio2023 has a header bu… Answer: Running DESeq on normalized data by James W….
Interaction terms in DESeq2
Hi, I am hoping this isn’t a stupid question as I am really lost here. I have extensively read the manual and other forum posts but am struggling to find a solution. I am using DESeq2 to analyse my data set but running into problems with an interaction term in…
KCouper/Liverpool K-means RNAseq Analysis November 2020
R3 VAR14 vs RBC no TNF k-means q0.05 1. Genelist Selection groupsName<-“R3_VAR14_kmeans_q0.05” countsTable<-read.delim(“RNAseq2019July_5.txt”, header = TRUE, sep = “\t”,check.names=FALSE,row.names=1) head(countsTable) AllGeneNames<-countsTable$Gene_Symbol #head(AllGeneNames) tempA<-countsTable topDEgenes <- which(tempA$padj_R3noTNF_var14_vs_RBC_0h<0.05&!is.na(tempA$padj_R3noTNF_var14_vs_RBC_0h))####find indexes listA<-tempA[ topDEgenes, ]$Gene_Symbol topDEgenes <- which(tempA$padj_R3noTNF_var14_vs_RBC_2h<0.05&!is.na(tempA$padj_R3noTNF_var14_vs_RBC_2h))####find indexes listB<-tempA[ topDEgenes, ]$Gene_Symbol topDEgenes <- which(tempA$padj_R3noTNF_var14_vs_RBC_6h<0.05&!is.na(tempA$padj_R3noTNF_var14_vs_RBC_6h))####find indexes listC<-tempA[ topDEgenes, ]$Gene_Symbol topDEgenes <- which(tempA$padj_R3noTNF_var14_vs_RBC_20h<0.05&!is.na(tempA$padj_R3noTNF_var14_vs_RBC_20h))####find indexes listD<-tempA[ topDEgenes,…
different results with and without releveling when providing contrast
Hello, I am conducting a differential expression analysis of RNA-seq data where I have samples from three treatment doses (control, low, high) at three developmental timepoints (W6, W8, GS45). Note these are not paired samples (i.e. different individuals sampled at each timepoints). I am interested in differentially expressed genes caused…
Solved Now write an R script with the contents shown below
Now write an R script with the contents shown below to import the Salmon alignments into `DESeq2` and perform differential expression analysis. As with the previous script, since I provide the complete script, look up each of the functions used and make sure you understand how they are used here….
How can I devise a method like DESeq2 to give statistics for paired multiple -omic datasets?
Hi, I am a PhD student seeking advice on how I can devise a method like DESeq2 to give statistics for paired multiple -omic datasets that are aligned with those produced on RNA-Seq count data by DESeq2 (one of the multi-omic datasets for paired samples), so that they can be…
Epigenetic factor associated gene serach
Hello Everyone, I have a set of genes that I got from my RNA seq experiment. This file looks like this : gene Symbol baseMean log2FoldChange lfcSE stat pvalue padj UP_DOWN ENSG00000001497 LAS1L 2669.490018 -1.11256317 0.276762731 63.9156085 4.35E-13 6.58E-12 DOWN ENSG00000002726 AOC1 39.71640451 -2.490906864 0.773899659 20.0813272 0.000481272 0.001685566 DOWN ENSG00000003096…
Exporting DEGs obtained from DESeq2
Hi, I am not sure if my “action” is against the rules of biostar, and if so, please cancel my post. My question has been discussed on bioconductor but I have realized that I may not get an an answer so I decided to open a new question and ask…
Deseq2 calculated log2foldchange not consistent with RPM in IP-small RNA-seq
Hello, I am using Deseq2 for differential expression analysis of my small RNA-seq data, specifically I want to know what small RNAs are bound by a protein, so I pulled down the protein and sequenced both IP and input. I got the reads assigned to each gene using featurecount and…
identify DEGs across all conditions and per specific conditions
Hi, I am analyzing a bulk-RNAseq and I want to analyse the dataset using Deseq2. I am very confused so apologies if it’s a stupid question. My dataset has 12 samples (3 per condition). the conditions are: treatment and control and 2 time points (0hr, 12hrs). So I wanted to…
DESeq2, results(dds, name = ) interpretation results
Hello together, hello Michael, I have an understanding problem using results(dds, …) I have RNAseq data and a grouping of two conditions with interaction of time with the defined reference levels, see code below. And run the code as below. coldata$Cond <- factor(coldata$Cond, levels = c(“A”,”B”)) coldata$Time <- factor(coldata$Time, levels…
GO ENRICHMENT ANALYSIS- DESEQ
I keep getting this Error message: preparing geneSet collections… –> Expected input gene ID: C9orf78,CNOT9,SPIDR,FAM9B,SDC1,CCDC182 Error in check_gene_id(geneList, geneSets) : –> No gene can be mapped…. Here is my code, i attempting to perform gene set enrichement analysis: GroupA <- results(dds, name = "Cancer_Type_healthy_vs_MESO") original_gene_list <- GroupA$log2FoldChange names(original_gene_list) <- GroupA$X…
Deseq2, enrichGO and ensembl ID
Deseq2, enrichGO and ensembl ID 1 @3cc02754 Last seen 5 hours ago United Kingdom Hi I used code initially in DESEQ2 dds=DESeqDataSet(se,design=~TRAIT) dds=DESeq(dds) res=results(dds) I currently have results from DESEq2 which looks like this: log2 fold change (MLE): TRAIT S vs N Wald test p-value: TRAIT S vs N DataFrame…
r – DEseq2 results, enrichGO and ENSEMBL ID not matching
I am trying to do DEG and then use enrichGO on the results. I used code initially in DESEQ2 dds=DESeqDataSet(se,design=~TRAIT) dds=DESeq(dds) res=results(dds) I currently have results from DESEq2 which looks like this: log2 fold change (MLE): TRAIT S vs N Wald test p-value: TRAIT S vs N DataFrame with 42800…
Is it okay to just average the log2FC values across different cell types in pseudobulk scRNA-seq data?
Is it okay to just average the log2FC values across different cell types in pseudobulk scRNA-seq data? 0 Hi! I downloaded a differential gene expression summary data table like this from brainSCOPE. All I need is “gene” and “log2FoldChange” column. However, each gene’s data is split into multiple different cell…
DESeq2 results – Annotating and exporting results
Hi, I am working with isoforms.results from RSEM analysis. I am trying to annotate my deseq results with symbol and entrez IDs, following the vignette master.bioconductor.org/packages/release/workflows/vignettes/rnaseqGene/inst/doc/rnaseqGene.html#annotating-and-exporting-results Unfortunately, I cannot export them as a csv file because the 2 elements I am adding are list. do you have any idea how…
Genome-wide analysis of circRNA regulation during spleen development of Chinese indigenous breed Meishan pigs | BMC Genomics
Overview of the sequencing information To explore the presence of circRNAs during spleen development, we assessed circRNAs expression in the spleen tissues of Meishan pigs at various developmental stage. We prepared and sequenced ribo-depleted total RNA-seq libraries, as shown in the flow chart (Fig. 1). Table S2 presents our rudimentary sequencing…
r – Resizing and adjusting text and plots dynamically in ggplot when generating EnhancedVolcano plots
I’m using the EnhancedVolcano package in R to generate plots, which utilizes ggplot2 under the hood. When I try to save the plots at various dimensions using ggsave(), I’ve noticed that the text and titles remain the same size, causing the main plot contents to shrink and become less visible…
Transcriptional variation in Babesia gibsoni (Wuhan isolate) between in vivo and in vitro cultures in blood stage | Parasites & Vectors
Morphological observation of continuous in vitro cultured B. gibsoni (Wuhan isolate) Babesia gibsoni was successfully cultured in vitro in 20% serum. After splitting, parasitemia reached 10% ± 1.5% on day 3 (Fig. 1A). Fig. 1 Changes in parasitemia and morphology of in vitro cultured B. gibsoni (Wuhan isolate). A Changes in parasitemia of…
Retrieve deseq2 normalised counts with only results file available
Retrieve deseq2 normalised counts with only results file available 0 I have several Deseq2 results file, so a csv file with ensembl name, baseMean , log2FoldChange, lfcSE, stat, pvalue, padj columns. Unfortunately, I don’t have the original count files. Is there a way to extract Deseq2 Normalised Counts from the…
To identify Differentially Expressed Genes and Unraveling Upregulated and Downregulated Genes using DESeq2 in R
Hello!! AIM: To obtain differentially expressed genes and then identifying upregulated and downregulated genes from my analysis, here are the steps I run on R(DESeq2): 1. counts <- read.delim(“counts.csv”, header = TRUE, row.names =1, sep = “,”, check.names=FALSE) 2. dim(counts) 3. colnames(counts) 4. colData <- read.delim(“colData.csv”, header= TRUE,sep = “,”)…
ComplexHeatmap with anno_barplot
I’ve been making various heatmaps for different gene sets and I added the log2FoldChange values as an extra column, but I need to leave it as a barplot, but I’ve been lost on how to put those log2FoldChange values to appear as bars. Any help is appreciated, thanks 😉 “`r…
Creating heatmaps with 2 Log Fold Changes in DESEQ2
I am a new learner of DESEQ2 and have seen similar posts on this forum but could not find clear answers. What would be the best way to create heatmaps specifically with 2 log fold change(base 2) in DESEQ2 or R in general? In DESEQ2, heatmaps are created based on…
Enhanced Volcano transcript label
Hey, thanks for the comments. There is a somewhat indirect way of doing this by modifying the transparency of the points via colAlpha. Take a look: library(airway) library(magrittr) data(“airway”) airway$dex %<>% relevel(“untrt”) library(“DESeq2”) dds <- DESeqDataSet(airway, design = ~cell + dex) dds <- DESeq(dds, betaPrior = FALSE) res1 <- results(dds,…
ranking the DeSeq2 results using t-values?
ranking the DeSeq2 results using t-values? 1 @theodoregeorgomanolis-7993 Last seen 6 hours ago Germany Hi all, this might be a bit complicated post I was trying to get a better than the log2fold change to rank my Deseq2 results, so that for example to get important genes ranked poth by…
DESeq2 lfcShrink function used in DiffBind package
DESeq2 lfcShrink function used in DiffBind package 1 @aaed3153 Last seen 9 hours ago United States Hello, We have basically three questions that revolve around the DESeq2 lfcShrink function which is used by DiffBind. We have Cut&Tag samples and want to conduct differential binding analysis. Our main objective is to…
Losing a lot of significant genes after removing outliers. Having cell type composition as covariates
Hi all, I am working with the RNA-seq data on humans (24patients-20controls). I used DESeq2 to find differentially expressed genes. here is the code that I used: It is corrected for cell-type composition (using cibersort and PCA on the estimated cell-type proportions) dds <- DESeqDataSetFromHTSeqCount(sampleTable=sampleTable, directory=folder, design=~Plate+RIN+Sex+Age+condition+PC2+PC1) #considering PC1,PC2 as…
DESeq2 how to extract expression values corresponding to log2FoldChange
DESeq2 how to extract expression values corresponding to log2FoldChange 1 @4e4abe33 Last seen 14 hours ago France Hello, I’m working with DESeq2, and when look my normalised mean expression in the Case vs Control the date doesn’t match with the log2 fold change. Example: gene baseMeanCase baseMeanControl log2FoldChange lfcSE stat…
Heatmap
Heatmap 0 Hi, I´m trying to do a heatmap with ggplot but when I´m going to represent my values (Excel) and I write the command: data_heatmap <- expand.grid(X=x, Y=y) head(data_heatmap) data_heatmap$Z<- runif(21, -3, 3) View(data_heatmap) ggplot(data_heatmap, aes(X, Y, fill= Z)) + geom_tile() + scale_fill_gradientn(colors = c(“red”, “white”, “blue”)) The values…
extreme high log2FoldChange in DESeq2 result
extreme high log2FoldChange in DESeq2 result 0 Hi all, I am working with the RNA-seq data on humans (24patients-20controls). I used DESeq2 to find differentially expressed genes. here is the code that I used: dds <- DESeqDataSetFromHTSeqCount(sampleTable=sampleTable, directory=folder, design=~Plate+RIN+Sex+Age+condition+PC2+PC1) #I used the principal component of combination of cell markers colData(dds)$condition…
DESeq2 for different design and normalized counts
DESeq2 for different design and normalized counts 0 Hi, I have normalized count data from RNA-seq protocol. The normalization steps include DESeq2 with design ~1. Unfortunately, I do not have the raw counts. Can I use those normalized counts for DESeq2 with different design? Thank you. DESeq2 log2foldchange • 84…
Remove duplicate genera in DEseq output?
Hi all, I successfully ran DESeq2, though I am looking for differential abundance of microbial genera, not gene expression. However, duplicate genera (assigned to different species in my taxa table) are listed out and numbered instead of collapsing. I would like to collapse duplicate genera. Here is what I ran…
Deferentially expressed gene with high log2foldchange by DESeq2; but not meaningful at the individual level
Hi all, I am working with the RNA-Seq data on human (24Cases-20 controls) to find differentially expressed genes. my RNA-Seq data is unstranded. Here is the comments that I used to align the fastq files: ls *_1P.fastq.gz | parallel –bar -j8 ‘R2=$(echo {} | sed s/_1/_2/) && out=$(echo {} |…
r – how to do a size changing by variable value in ggplot?
In geom_tile the size aesthetic only controls the size of the line (now replaced by linewidth in newer versions of ggplot2). The tiles themselves cannot really change size as their filling of the whole space is part of how that geom represents data. You can use geom_shape with the squares…
DESeq2 with multiple factors
DESeq2 with multiple factors 0 I have 18 samples. These samples have 2 different factors with one factor(Infestation) having 2 levels (as control vs affected) & another factor(timepoint) having 3 levels(as 24h, 48h,96h) . Also the last factor (timepoint) have 3 biological replicates. While performing: counts <- read.delim(“counts.csv”, header =…
Can I consider these values as differentially expressed?
Hello! I’m needing some help from the more experienced ones! n_n’ I’m doing a transcriptome expression comparison using DESeq2 and I would like to be sure I’m using the right parameters. This doubt came after seeing that a gene increased the expression between different sampling points but was not included…
non-numeric argument to binary operator
My Volcano plot is working fine for the argument on one shape for all as below; EnhancedVolcano(res_df, lab = res_df$gene_sym, x = ‘log2FoldChange’, y = ‘pvalue’, title=”MSC Cellular RNA vs MSC Exosome”, pCutoff = 0.05, FCcutoff = 1, pointSize = 3.0, labSize = 6.0, shape = 23, colAlpha = 1)…
data file link: | Chegg.com
data file link: drive.google.com/file/d/1Odr12yDiUwI02-BfaXrHehKBM1uMW_1N/view?usp=share_link Step 1 (5pts) Load the file GSE124548.raw.txt into R and create a new dataframe with just the columns with the raw counts for healthy (HC) and CF patients before treatment (Base) and call it readcount. Use the first column (EntrezID) in the original file as the…
Inverted log2foldchange in study?
Hi guys, I’m relatively new to any type of analytics work and Rstudio. However, when inputting my TCGA data and doing all of the basic steps, my volcano plot and log2foldchange values seem to all be the opposite of what was expected. I understand perhaps this could just be the…
Identification of cancer-related genes FGFR2 and CEBPB in choledochal cyst via RNA sequencing of patient-derived liver organoids
Background: Choledochal cysts (CC) are congenital bile duct anomalies with 6-30% risk for developing bile duct cancer. However, the molecular mechanisms underlying cancer risk of CC are unknown. We sought to identify the gene expression changes underlying the cancer risk of CC patients. Methods: Liver organoids (n = 51) were…
Meaning of stat values in DESeq2 result
Meaning of stat values in DESeq2 result 1 Hello, experts. I’m curious what will be the meaning of stat values in the DESeq2 result. For example, if geneA has a stat value for 50, and geneB has a stat value for 20, what is the overall comparison meaning of these…
different package different padjusted and qvalue
Hi! Apologies for the stupid question! but I think I am doing something wrong but i do not understand what. I would like to do ORA analysis on bulk-RNAseq dataset so I tried both clusterProfiler and also genekitr.` However, despite getting the same terms, but I have different p-adjusted value…
boxplot issue
I am working DESeq2 i wanted i proper boxplot but i don’t understand what wrong i am doing and i wanted to know how to properly plot the x and y axis what parameters should be taken for boxplot . here’s my code: library(“DESeq2”) library(“ggplot2”) counts<-read.delim(“PC_1.csv”,header = TRUE, row.names =…
Extract/include transcript list from GAGE analysis result
Extract/include transcript list from GAGE analysis result 0 Hi everyone, I’m using GAGE following Deseq2 analysis on my RNAseq data. This means, the analysis is being done on log2FC. I can’t see any list of transcripts related to GO terms/pathways that are being enriched. Something like the usual tools of…
DESeq2 for time series and control-treated groups
DESeq2 for time series and control-treated groups 0 Hi, I am new at DESeq2 and trying to desing matrix to get DEGs. Basically, I need number of DEGs based on compare control/treated groups each time points. I tried dds <- DESeqDataSetFromMatrix(countData = round(counts_data), colData = colData, design = ~time +…
Variation in number of DEGs on LFC shrinkage
Hi, I am using DeSeq2 to estimate the DEGs across a dataset of 6 samples with 3 samples each in 2 conditions – patient versus control. The parameters for the DEGs are as follows: |Log2FC|<2, P-value<0.05 I ran DeSeq on the datasets through 2 ways – (i) Without the LFC…
Overlay points with EnhancedVolcano R
Hi everyone, I would like to colour a specific population of genes in a volcano plot using enhanced volcano function. The thing is they are crowded in the rest of the non-significant genes. Would like to overlay them to be visible. Here is the code used keyvals.colour <- ifelse( res_table_tb_sex$ygenes…
no gene can be mapped (RNAseq analysis)
gseKEGG – no gene can be mapped (RNAseq analysis) 0 Hi all, I have been trying to extract the GSEA results from a list of genes after RNAseq analysis. It looks like my gseKEGG function is giving me problems. I am unable to generate a list of KEGG terms, it…
How to employ statistical approaches to identify differentially expressed genes (DEG)
Identifying differentially expressed genes (DEGs) is an important task in genetic research because it allows researchers to identify genes that are associated with a particular phenotype or condition of interest.DEGs are genes that show significant differences in expression levels between two or more groups, such as a disease group and…
Merge common elements in R
Hello everyone, I have a list of differential genes (list1) and another list where I have the ID of the genes and the name (list2) and I want to name the genes from list1 taking the name from list2, how can I do this? I will appreciate your support, thanks….
DGE/DEG Analysis for comparing multiple cell lines
Hello community, I’m relatively new to DGE/DEG analysis using RNA-Seq data, for which I’ve seen that DESeq2 is one of the go-tos for differential gene analysis. I am a bit confused about the list of genes I am obtaining and which type of normalization methods are best to use (variance…
DE Analysis on cells from a patient derived mouse xenograft with high levels of mouse count “contamination”
I am performing a differential expression analysis for collaborators. The overall biological design from my collaborators is as follows: 1) Received patient sample. 2) Amplified patient sample using patient derived xenograft (PDX) in a mouse host. 3) Extracted cells from mouse and enriched for human cells by positive selection using…
tximeta accessing gene symbols after addIds()
Hi, I’m sure I’m missing something obvious, but is there an easy way to access the gene symbols after adding them with the addIds() function in tximeta? gse <- addIds(gse, “SYMBOL”, gene=TRUE) mcols(gse) DataFrame with 60240 rows and 3 columns gene_id <character> ENSG00000000003.15 ENSG00000000003.15 ENSG00000000005.6 ENSG00000000005.6 ENSG00000000419.12 ENSG00000000419.12 ENSG00000000457.14 ENSG00000000457.14…
Help for RNA sequencing analysis with DESEQ2
Help for RNA sequencing analysis with DESEQ2 0 @80066fff Last seen 1 day ago Canada Hello everyone, I’m currently trying to perform DESEQ2 with my own RNA sequencing results. I performed whole transcriptome RNA sequencing and the subsequent analysis I performed via galaxy. I performed first fastp on the samples,…
Bioinformatics Analysis of the Key Genes and Pathways in Multiple Myel
Introduction Multiple myeloma is a B-cell malignancy characterized by the malignant proliferation of clonal plasma cells in bone marrow,1 It is the second most common hematological malignancy after lymphoma.2 The age-standardised rate (ASR) of multiple myeloma incidence was 1·78 (95% UI 1·69-1·87) per 100 000 people globally and mortality was…
Log2FC values slightly higher in some genes after DESeq2 shrinkage
Hi, I have a question about DESeq2 LFCshrinkage: Is it possible that some genes have a slightly higher LFC after shrinkage? It happened during my RNAseq DE analysis, I have very deeply sequenced samples with large base means. I tried visualizing using MAplot check, and it looks fine. I’m mainly…
between results of RNAseq and absence/presence of Type3 Secretion System
Correlation between two different datasets: between results of RNAseq and absence/presence of Type3 Secretion System 1 Dear All, I have a “How would you solve” kind of question. I have two sets of tables : 1. Log2FoldChange table and 2. Effectors Table. Firstly, the Log2FoldChange table was obtained by performing…
Seeking help in multifactor differential gene expression analysis using DESeq2. Can not get significance difference while comparing 3 time factor (0,2 and 4h) with 3 group (1 control, 2 treated)
Dear Experts, I have RNAseq data from 6 different plant samples (2 control, 2 Sensitive, and 2 tolerance), and different location of one species. I am trying to see the effect of the different groups at different time points, but after going through all the posts and vignettes I am…
InteractiveComplexHeatmap on DESeq2 object with more than 2 groups
InteractiveComplexHeatmap on DESeq2 object with more than 2 groups 1 Hello all, I’m writing with the hope someone can clarify a doubt I have about the differential heatmap generated by the package InteractiveComplexHeatmap via the simple command interactivate(dds). I read the package documentation at bioconductor.org/packages/release/bioc/html/InteractiveComplexHeatmap.html, but I couldn’t find the…
Genes looking differential abundant are not accoring to DESeq2
Genes looking differential abundant are not accoring to DESeq2 0 I have a metagenomic dataset crossing three time points from which I have mined CAZymes and am using DESeq2 to identify differentially abundant CAZymes from using trimmed mean depth generated my CoverM (very similar to Q2Q3 contig coverage). From this…
Finding significant up-regulated and down-regulated genes given a log2FoldChange threshold
I have a result from which I want to get significant genes (pvalue-threshold=0.1, log2FoldChange threshold 1.5), and divide them into up-regulated and down-regulated. From the vignette, I found the following on down-regulation and up-regulation: subset the results table to these genes and then sort it by the log2 fold change…
r – Filtering data and labeling them based on threshold issue
I’m trying to set a threshold with different criteria to label data points into categories So my code is this C1_C2 = read.csv(“C1_C2_contrast.txt”) diff_C1_C2 <- C1_C2 %>% mutate(UP_DOWN = case_when( # padj < 0.05 & log2FoldChange > 1.5 & log2FoldChange < 2.5~ “UP”, baseMean > 50 & pvalue < 0.05…
Fatty infiltration after rotator cuff tear
Introduction Rotator cuff tear (RCT) is a common shoulder disorder causing shoulder pain and disability. The prevalence of full-thickness RCT is 20.7% in the general population, and increased with age.1 Rotator cuff play essential roles in shoulder function and the treatment of proximal humeral fractures.2,3 It is important to repair…
How to do functional analysis on differentially expressed gene list from RNA-seq data?
How to do functional analysis on differentially expressed gene list from RNA-seq data? 0 @06f08eeb Last seen 1 day ago Canada Hi all, I am a complete beginner in terms of bioinformatics analysis and I am hoping to complete some functional analysis on some differentially expressed gene lists of some…
Strangely too low P-value and Adjusted P-value(FDR) DESeq2 and edgeR
My data is experimental data that has been overexpressed for a specific gene. Data samples are divided into 3 groups according to the over-expression time and each group has 3 samples. (total 9 samples) I conducted DGE analysis on the control group and one case group with DESeq2. cts <-…
Change log2FoldChange range – plotMA
You can use base R graphics to make these plots. The data is sitting there in columns of the res object, so you can filter it directly, and use boolean vectors to pick out the things you need: # make sure there are no NA values sum(is.na(res$log2FoldChange)) # choose some…
sigh of log2FC values in DESeq2
sigh of log2FC values in DESeq2 0 @194b0276 Last seen 10 hours ago United States Can someone explain to me how is the sign of log2FoldChange is set in the results of DEseq2? I was pretty sure that it is calculated as log2(Counts_treatment/Count_reference), where reference is determined alphabetically (unless specified…
highlighting specific genes (from a user-supplied list) in a Volcano plot in R
highlighting specific genes (from a user-supplied list) in a Volcano plot in R 1 I’ve generated a volcano plot using DeSeq2 results and would like to specifically highlight a subset of genes by providing a list of gene IDs Dataset$condition <- relevel(Dataset$condition, “Ctrl”) res <- lfcShrink(DatasetProcessed, contrast=c(“condition”,”Treat”,”Ctrl”)) with(res, plot(log2FoldChange, -log10(pvalue),…
TPM value from DESE2 and significant filterig isssue
TPM value from DESE2 and significant filterig isssue 0 The code 10101 res_ddsDE_new has 36,000 rows. When I am using subset(res_ddsDE_new, padj < 0.05 & abs(log2FoldChange) > 1) res_ddsDE_new baseMean log2FoldChange <numeric> <numeric> DDX11L1 1.779144 -1.4955939 WASH7P 152.518293 -0.0505911 MIR6859-1 20.653876 0.5689275 MIR1302-2HG 0.255387 -1.9691031 FAM138A 0.353478 0.1574042 Then I…
Understanding the output of Negative Binomial in DESeq2
Are zero is a default option of log2fold change to be considered as up and down? Yes, by default, the null hypothesis is that the log2FoldChange is zero. Does this mean that there are seven genes in total that will have significant adjusted p-value? Yes, you are interpreting your results…
DESeq2 log2FC and p-value filtering
DESeq2 log2FC and p-value filtering 1 Dear all, I want to filter DE genes by log2FC > 2/-2 and p-value 0.0001. Why this two versions of lof2FC and p-value filtering differ in up- and down-regulated genes numbers (from 800 to 1500) and which one is the right one ? 1….
Correct way to make multiple comparisons on DESeq2?
I have a project where I have done RNA-seq (paired-end sequencing on Illumina HiSeq) of a worm at different days of development i.e. Ages 0-12. For each age, I have sequenced 3 replicate specimens. I’m new to DESeq2 and I was wondering if what I did below is correct. library(DESeq2)…
Outliers on DESEq2 Results
I have an RNAseq dataset, where one of the genes I intend to analyze has hundreds of counts ranging from 10 to 12, with a few counts > 9000. I process this data in Deseq2 and get that the gene is differentially expressed across several samples of interest. What can…
Need help to remove NA values from data frame
Need help to remove NA values from data frame 2 I have this data frame : and I want to remove those rows which contain NA values from the log2fold change column How can I do this through R? DeSEQ2 R • 256 views Hi Anas, If your data frame…
Volcano plot not centered at origin
I am making a volcano plot for my phosphoproteomics data and seemingly the data are not centered around the origin. The reason that I know there is something wrong is because before my code was producing the correct volcano plot, and now it is shifted. What are some reasons why…
Handle inflated log2FC while using interaction term in DESeq2
Hi guys, I’m working with a 8 samples experiment (lower than 3x replicates, I know..) with a design like > colData(dds) DataFrame with 8 rows and 2 columns condition traitment <factor> <factor> 4200-JS-1 norm ctrl 4200-JS-2 norm ctrl 4200-JS-3 norm trt 4200-JS-4 norm trt 4200-JS-5 hyper ctrl 4200-JS-6 hyper ctrl…
Is it a good idea to plot a volcano plot starting at position x =1
Is it a good idea to plot a volcano plot starting at position x =1 1 Hello everyone! I was wandering if it makes sense to start a volcano plot (x=log2FoldChange) starting at position one, so basically discarding all values below 1 and above -1 leaving only values outside this…
Circos plot with logfold change RNA seq data
I am new to circos plot analysis and have been trying to use the cyclize package. I want to display mRNA differential gene expression data based on data analyses of 8 libraries and links between their respective target genes. The dataset I am working with looks like this geneid baseMean…
lfcShrink probelm in many 0 count genes RNA-seq data
Hi, Dr love. I post a question about weird MAplot or volcano plot of DESeq2 diff result and also in biostar. ATpoint give a useful answer about too many 0 count genes and prefiltering. It seems that too many 0 count genes makes lfc shrink have a probelm. And I…
weird MAplot or volcano plot of DESeq2 diff result
Hi, every one. I find a werid MAplot or volcano plot of DESeq reuslt. I am wondering whether you can give me some advice. This diff result is from two cell type bulk RNA-seq. I use two specific marker to get these two cell type using Flow cytometer. I alreadly…
same padj for all the genes after DEseq analysis
Hi everyone, I have done a pair comparison with DEseq2 to find differentially expressed genes between two samples with 6 replicates, for the DEseq2 result, i got exactly same padj value for all the genes and it is not significant, is this normal ? I don’t think the padj should…