Questions tagged [single-cell]
Questions about techniques in which the genetic material of a single cell is analysed individually
164 questions
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BCL files half the size of fastq files (10x chromium single cell)
I was just curious about this. The BCL files (~100GB) we have for a single cell experiment (10x chromium), are less than half the size of the produced fastq files (240GB, gzipped). This is excluding ...
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How to count the number and proportion of different barcode combinations in DNA sequencing results
This question was also asked on Biostars
We used a new single-cell sequencing method for sequencing, and now we have encountered the following problems when analyzing the data. We designed three ...
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Demultiplex double hashed cells using Seurat (R)
I am using Seurat to demultiplex my single-cell RNA data, which also has HTO data for hashing. Each cell is hashed once based off it's origin ie organ1, and then hashed based off the cell type ie cell ...
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10X Cellranger count - Unable to distinguish between [SC5P-R2, SC3Pv2] chemistries based on the R2 read mapping for sample GEX in
Recently ran three separate single-cell experiments and sequenced them all using the 10X platform. Each of the experiments involved GEX, FBC and VDJ. With the resulting BCL files, csv files with info ...
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Cell to Cell communication detection tools for wide range of species
I am starting to get interested in protein interactome analysis from single cell/nucleus datasets.
I found three tools that might be interesting, which are ...
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How can I quantitatively evaluate which UMAP is best in terms of clustering & embedding?
This question was also asked on Reddit
I am new to sc-rna analysis, I have the dataset that I am trying to find out the best UMAP, experimented on trying out different values of the parameters as in (<...
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Should I use cellranger count or multi for biolegend hashtags?
I have a set of fastq files. One of them is for the gene expression data and the other is feature barcode data which contain hashtag information.
I was told to use cellranger count for feature barcode ...
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Find up/downregulated gene lists across scRNA-seq samples
I have two separate scRNA-seq datasets that are different batches. I want to get genelists of up/downregulation of one cluster in one dataset compared to another cluster in the other dataset.
I ...
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Practical challenges for microarray analysis and single cell
currently, I am learning single-cell sequencing and microarray analysis. I am searching for somewhere (or any alternative way)to practice and get hands-on with these techniques. please help me with ...
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How to perform meaningful Gene set erichment analysis or otherwise find broader themes/functions in different cell populations?
(For context: I'm somewhat new to bioinformatic analyses but am mostly comfortable with R)
I have identified transcriptomically and morphologically different cancer cell populations within a patient (...
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Convert ensemble genes to gene names in my sigle cell signature matrix annotated with seurat in R?
I have single cell data which I want to annotate with the seurat package, so I ran this code in R:
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Identification of metabolites through spectral library search
I am very new to single-cell metabolomics and today I want to jump in this area. I have been reading many review papers related to identification of metabolites at single-cell resolution from spectral ...
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scRNA: What are good dimensionality reduction/clustering parameters to get biologically plausible groupings?
I've got a moderately large set of PBMCs, over 1M cells. That means I can't easily do a grid search of dimensionality reduction/clustering parameters/methods. Some examples results I'm getting with ...
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Parsing gene names to detect organism from h5ad input
I have one function that is for reading .h5ad files
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Using CSV files continaing scRNA-seq count data from GEO [closed]
I'm completely new to scRNA-seq analysis (and to most of Bioinformatics as well) so I apologize if this was asked a million times before.
I am trying to get the single cell expression data of lymph ...
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Loading scATAC-seq data containing matrix.mtx, fragments.tsv and barcodes.tsv files from GEO into R?
I want to analyse the single-cell ATAC-seq data from 10X downloaded from GSE158398. The files attached to this dataset are matrix.mtx.gz, ...
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What trajectory analysis method allows to set the form of the trajectory?
I have single cell data from two samples : normal and pathological and
I would like to track the progression of the cells (and find the genes that drive it) from normal to pathological.
Upon first ...
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Cell Ranger returns too few cells!
I recently performed single cell RNA seq using core services and the web summary output from 10x that they have given shows that there are very few cells detected (100-300) cells per sample. Once this ...
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shiny app for single cell cluster export issue
I'm trying to create a shiny app for single cell data visualization and then label cells with genes till this step it works, the final step when I try to export cluster it is not exporting the desired ...
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Plotting infercnv results
I'm working with matched single cell data, where we have treated and untreated samples for the same patient. I ran CNV analysis using the infercnv package.
I've ...
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h5ad file format filter
I'm trying to do simple filter for single data that is stored in h5ad file format using this
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Find Lipidic Mediator receptors
I want to study the ligand-receptor interaction and I am searching for lipidic ligands( mediators), I have found 16 so far. Does anyone know any database where I could find more lipidic ligands. I was ...
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Identifying somatic mutations in cell lines
I would like to identify the somatic mutations present in a cell line and characterise the genes that are potentially affected by those mutations. For example, are there oncogenes mutated in a ...
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How to calculate cell type percentage in every sample
I have a Seurat object (metadata) with the single R samples consisting of cell types and sample types columns. I am trying to make a table that has a sample and percentage of cell types for each ...
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How to make a UMAP for single cell data and color cells by average expression of a list of genes in scanpy?
I would like to make a UMAP where the cells are colored by the average expression of the bulk signature genes but I am not confident that I did it correctly. I would like to use scanpy for it.
I did ...
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Can I use palantir for trajectory analysis in a a merged object?
I was asked to use palantir for trajectory analysis. I have 4 samples that I have preprocessed using scanpy. Can I run the palantir analysis on the merged object or do I need to unmerge them?
Thank ...
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How to do pathway analysis after scanpy for single cell data after DE analysis?
I have done DE analysis using SCANpy on my single cell data and I have compared each cluster versus all the other clusters. One cluster seems particularly interesting so I wanted to do pathway ...
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How to identify a low proportion cell subpopulation in the single-cell RNA-seq data?
The cell subpopulation that I am interested in only accounts for around 1.2% of the total cells. I have previous FACS experiments that sort out the subpopulation from the samples (using markers CD166+ ...
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pyScenic CLI for ctx is giving error: Not a single module loaded
Hi I ran pyScenic's ctx via the command in command prompt:
...
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How many percents explained variance by the first 50 principal components of PCA analysis is suitable for downstream `KNN` and `umap`?
In single-cell RNA sequencing analysis, I am wondering how many percents explained variance by the first 50 principal components of PCA analysis is suitable for downstream ...
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How to concatenate 2 `mdata` of `muon` package?
I used muon(https://github.com/scverse/muon) for single cell Multimodal omics analysis.
How to concatenate 2 mdata from ...
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Is there a package that can filter the `raw_feature_bc_matrix` of 10X's `cellranger multi` pipeline output?
There is no filtered_feature_bc_matrix in 10X Genomics' cellranger multi pipeline output when multiplexing analysis was ...
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Best method to compare differentially expressed genes between 2 single cell clusters for GSEA
I want to get a ranked gene list for GSEA analysis from single cell data, which needs a group differential expression statistic for ranking.
Which is the best method to compare the differently ...
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How to preserve colour assignments in matplotlib when a category is empty?
Some of my samples have empty clusters. I want to keep the same color for each cluster in different plots.
How do I get sc.pl.umap to keep the same colors for each ...
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FlowSOM multi-step clustering
There are a few different commonly used clustering algorithms within the single-cell space, although Leiden seems to be the top choice these days. FlowSOM is a classic package for analyzing flow ...
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How to extend the x axis in Dimplot Seurat
I have performed a Seurat PCA via Dimplot.
How do I extend the x axis?
As you can see in my figure the double x axes overlap.
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Definition of genotype in demuxlet
I am reading the Online methods of the demuxlet paper.
The genotype $g$ is taken from the set $\{0, 1, 2\}$, defined as "the true genotype of the sample corresponding to $c$-th droplet at $v$-th ...
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Error in dimnamesGets(x, value) when trying to read data using Seurat package
This question has also been asked on Biostars
I am trying to create a Seurat object using the package "Seurat". When I am reading my raw files using the function ...
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extracting raw gene counts based on the cluster generated by seurat
I'm new to single cell sequencing analysis. I have made different clusters using seurat. Now I would like to extract the raw gene counts based on the cluster generated by seurat.
i have provided my R ...
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Identifying highly variable genes in scRNA-seq: Seurat vs M3Drop
Following a single-cell RNA-seq workshop, I created a Seurat object (my_data), normalized the data, and then tried to identify highly variable genes using two ...
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snRNA-seq Demultiplexing before or after QC
I'm currently analyzing snRNA-seq data and have to demultiplex them using HTODemux in Seurat.
The question is : should I do my QC filters on my droplets before or after the demultiplexing ?
From what ...
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how to change the pseudo time trajectory direction?
Monocle is a comprehensive bioinformatics package which has the function to run the pseudo-time trajectory and changes in direction.
I wish to replicate this analysis. Like in the figure I would like ...
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How can to validate the presence of a certain type of cells in a single cell dataset?
I have a single cell dataset which I consider to be a reference , let say, for an human organ, I have identified some new clusters that correspond to cell types. I also have WGCNA modules that ...
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Cellranger results have too many cells
I have the results of cellranger analysis for single-nucleus RNA seq data that was done by someone else. So I do not know which parameters were used for cellranger. In the results, there are 77000 ...
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How to convert 10x matrix.mtx.gz files to hdf5 format?
I have a bunch of folders containing barcodes.tsv.gz, features.tsv.gz, and matrix.mtx.gz from the Cellranger Count output for a single-cell dataset that was sent to me from another lab. I need the .h5 ...
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How can I get data of single cell RNA sequence with raw count?
I am using dataset GSE85241, but I can't find the read counts of the dataset. It only provides with RPKM values.
How can I find the read counts of a dataset?
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Does the number of RNA reads per cell obtained from the 10X scRNA experiment depend on amount of mRNA in given cell?
As we know, the amount of RNA reads per cell obtained from 10X scRNA experiment vary between cells. I wonder if this is effect of technical issues or does the number of RNA reads per cell obtained ...
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What genes are driving a certain pathway
I've currently run pathway analysis on a set of genes. I'm now trying to figure out how to see which genes are driving each pathway.
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Subsetting with harmony
Currently merged two Seurat objects together and then ran Harmony for batch correction. Now I want to subset out a cell type of interest do I re run harmony or just do the standard PCA?
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