Questions tagged [demultiplexing]
The demultiplexing tag has no usage guidance.
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Running cellranger on scRNASeq data with feature barcoding (10x + antibody capture)
I can't seem to find a clear answer to this question, so here it goes:
I have sequenced scRNASeq + scVDJSeq (TCR) data, which has been sequenced using feature barcoding from 10x genomics, via antibody ...
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STACKS: process_radtags only reads 1 input file and returns >90% barcode not found drops
I am trying to demultiplex some paired-end ddradseq data and am running into an issue with STACKS in that the program only seems to read 1 of my files for input (there are 2) and also results in over ...
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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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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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Why is bcl2fastq2 taking so long to calculate stats?
Our lab has been using bcl2fastq v2.20.0.422 to demultiplex RNA-seq data sequenced on an Illumina Novaseq machine on a beefy EC2 instance and we've run into the strange problem: namely that while ...
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Get frequency of index sequences of fastq file
I recently did some sequencing on a MiniSeq and unexpectedly the Undetermined_S0_L001_R1_001.fastq.gz file contained quite a lot of data (about 40% of total). I ...
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Demultiplexing FASTQ file without index information
I am trying to understand how data that was uploaded to SRA (https://www.ncbi.nlm.nih.gov/sra?LinkName=biosample_sra&from_uid=4510743) can be analyzed with the assumption that the FASTQ file ...
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Error processing run file with bcl2fastq - Unable to open '.../RunInfo.xml' file for reading
I'm attempting to demux a nextseq run which I've successfully done several times in the past on other runs. I uploaded the run from my PC to the server (Ubuntu 18.04.4 LTS) without issue. Then I ran:
<...
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2
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cellranger mkfastq with full path to --id flag
I have always used cellranger mkfastq to demultiplex 10x genomics runs manually, though recently the commands to do so have been incorporated into a script that ...
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How to demultiplex a mix of single-indexed and dual-indexed samples
The problem
If I have a sample sheet that contains both single-indexed and dual-indexed samples, I can split it up into two sample sheets and then run bcl2fastq on each one. However, when doing this, ...
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Demultiplex nanopore reads with custom barcodes
We have a problem trying to demultiplex MinION sequences with custom barcodes. Do you have any software recommendations we can try for demultiplexing or how to demultiplex these custom barcodes with ...
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Fastest way to demultiplex bam file based on field
I have a bam file with aligned reads from multiple experiments. For each experiment I have a portion of the sequence identifier, thus for example
...
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What is the index fastq file (sample_I*.fastq.gz) generated when demultiplexing Illumina paired-end runs?
What is the index fastq file that comes with some Illumina sequencing datasets? (The samplename_I*.fastq.gz file.)
For example, I recently received some 10X ...
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Demultiplexing and preprocessing for custom single-nucleus Drop-seq data
I am trying to reproduce the preprocessing of paired-end sequencing reads in Lake et al. (ref. 1).
First, paired-end reads were filtered out if read 1 had more than four non-T bases in the last ten ...