Category: logFC

RNAseq Viz

RNAseq Viz 0 Hi All, I’m really excited to have my first RNAseq expt. results back. However, I’m trying to find the best pipeline to work with in R that best matches my coding experience (little to none). I came across Searchlight2 which seems perfect as it is automated and…

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KEGG passway analysis after Seurat analysis

KEGG passway analysis after Seurat analysis 0 Hello, everyone. I am so sorry for this amateur question. I have a scRNA-seq data and want to compare and visualize the expression levels across genes on a cluster by using clusterprofiler package (passway analysis), but I think the average expression levels of…

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Do we need to filter out the 0 counts sgRNA in CRISPR analysis?

Do we need to filter out the 0 counts sgRNA in CRISPR analysis? 0 Hi, I’m doing CRISPR analysis and there is a question, when we perform CRISPR analysis, is there a step to remove low count sgRNAs from control and/or treatment samples? The reason of this question is that,…

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GOChord plot problem when using the chor_dat function to create a matrix

GOChord plot problem when using the chor_dat function to create a matrix 0 Hello All, I am trying to create a GOChord plot for circular visualization of the results of gene- annotation enrichment analysis. I created all the data frames myself and cross-checked the types of every column in each…

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Comparative cellular analysis of motor cortex in human, marmoset and mouse

Statistics and reproducibility For multiplex fluorescent in situ hybridization (FISH) and immunofluorescence staining experiments, each ISH probe combination was repeated with similar results on at least two separate individuals per species, and on at least two sections per individual. The experiments were not randomized and the investigators were not blinded…

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Statistical technique for mutation to gene expression link

Statistical technique for mutation to gene expression link 0 I grouped the samples into p53 wildtype and p53 mutated – for approximately 1000 individuals. I have gene expression data (logFC) of each individual present in both mutated and non-mutated groups. Now my aim is to identify the genes that are…

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Comparing two RNA-seq data sets mouse and human

Comparing two RNA-seq data sets mouse and human 0 Hi I have a question. We have two RNA-Seq data sets one from mouse and one from human for our gene modification, and we wanted to know how similar they are from each other. I know i can convert the mouse…

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How to work out Z score for heatmaps for RNA seq dataset

How to work out Z score for heatmaps for RNA seq dataset 2 I am trying to generate some heatmaps for my RNA seq dataset but struggling to work out how to calculate the z score. Can anyone give me any pointers on how to calculate please. I have; Gene_DE.txt…

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rs2507799 was found to be linked with increased risk for IS

Introduction Ischemic stroke (IS) is caused by the sudden loss of blood circulation to an area of the brain that causes injury to neurological function and represents a major cause of global disability and mortality.1 IS is known to be a heterogeneous and multifactorial disease. Genetic factors, particularly those involving…

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Why there are same name with different value in my microarray result?

Why there are same name with different value in my microarray result? 0 I have done one color agilent microarray. I am processing my data, but my data showed one miRNA present in more than one place with different value. Could someone please tell me what is the reason for…

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Introducing NA by coercion error message when running pathfindR

Since the weekend I’ve been getting an “NAs introduced by coercion” error message when i run the pathfindR package. An example of some of my data is Gene.symbol <- c(“ACAA2”, “ACADVL”, “ACAT1”, “ACOT9”, “ACOX1”, “ADH5”, “AKR1A1”) logFC <- c(“3.3”, “3.9”, “1.5”, “1.7”, “2.4”, “1.9”, “1.7”) adj.P.Val <- c(“0.02”, “0.03”, “0.02”,…

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Cutoff value for shrunken logFC

Cutoff value for shrunken logFC 0 Hello Is there any rule of thumb for filtering shrunken logFC, similar to |logFC|>1.5 for raw results? Or perhaps one should find the cutoff purely based on the distribution of shrunken logFC? I think some standardized value is needed though, e.g. from the perspective…

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relationship between DE-genes and DE-trascripts in deseq2

relationship between DE-genes and DE-trascripts in deseq2 1 Hello everybody, I ran a DE –RNAseq project by star-stringtie2- deseq2 pipline. The prepDE.py was used for generation of transcript matrix and gene matrix. Deseq2 were performed at both gene and transcript levels. At the gene level, 838 DE-genes were identified with…

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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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How to transform the deg gene list from seurat to a gene list input to clusterProfiler compareCluster ?

Sorry for lateness, I wanted to do something similar. This is what I did for reference: Using a Seurat generated gene list for input into ClusterProfiler to see the GO or KEGG terms per cluster. I’ll keep the meat and potatoes of the Seurat vignette in this tutorial: library(dplyr) library(Seurat)…

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Use Spike-Ins or TMM-normalization

Use Spike-Ins or TMM-normalization 1 Hi all, Sorry for all my questions lately, but as a novice which has to figure out how to analyse QuantSeq data, this forum has been a great and indispensible help for me. I’m doing a human transcriptomics analysis where we have QuantSeq data for…

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Calculate fold change in edgeR with one sample per condition

Hi, We have run a pilot RNA-Seq study with one sample per condition, this is just a test run. I understand there is no valid statistical test in this case, however just curious to obtain differential expression through edgeR package in R assuming dispersion = 0.4 for the human data….

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How to convert log2 scale RNA-Seq expression data to linear scale data

How to convert log2 scale RNA-Seq expression data to linear scale data 0 Hi, We have run a pilot RNA-Seq study and I used edgeR package to obtain differential expression results. The results output a gene column along with the logCPM, logFC and p-value column. I have a question regarding…

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Agilent-016436 Human miRNA Microarray 1.0

Upon request, a quick tutorial for processing the Agilent micro-RNA (miRNA) microarray data of GSE28955. The raw TXT files are contained in: ftp.ncbi.nlm.nih.gov/geo/series/GSE28nnn/GSE28955/suppl/GSE28955_RAW.tar Download this TAR file Unpack it [the TAR file] Unzip the txt.gz files Store these [txt files] in a directory raw/ Then, create a tab-delimited file, targets.txt,…

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TPM to logFC and pvalues

Hi, I assume you have to find differential expression between two cell lines (Cx and Dx groups). Since you need logFC and Pvalue, this R code can work. And you can use obtained matrix (mysample) to calculate FDR of your interest. mysample <- read.table(“./mymatrix.csv”, sep=”,”, header=TRUE) for(i in 2:nrow(mysample)) {…

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Power calculation for microarray data

Power calculation for microarray data 0 I have an initial sample of 228 patients from a microarray study. Recently I have obtained a new set of labels specifying different condition types for only 69 out of the 228 patients. I wanted to run a DEG analysis on this set of…

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Differential expression analysis of TCGA data based on tumor staging

Hi everyone I wanted to analyze TCGA-BRCA data for identifying DEGs in different TNM stages (I to IV) between Normal and Tumor. How to change the following code to get the DEGs based on the staging? CancerProject <- “TCGA-BRCA” DataDirectory <- paste0(“../GDC/”,gsub(“-“,”_”,CancerProject)) FileNameData <- paste0(DataDirectory, “_”,”HTSeq_Counts”,”.rda”) query <- GDCquery(project =…

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%% error in Rstudio

%% error in Rstudio 1 dc.markers %>% group_by(cluster) %>% top_n(2, wt = avg_logFC) the above code is giving error even after using dplyr and matrix libraries in seurat analysis in rstudio error : Error: Problem with filter() input ..1. i Input ..1 is top_n_rank(2, avg_logFC). x object ‘avg_logFC’ not found…

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