Tag: RSEM

Collapsing Transcript Count Matrix to Gene Count Matrix

Collapsing Transcript Count Matrix to Gene Count Matrix 0 Hi all, My colleague is trying to do some DGE. He got hold of a transcript count matrix from another colleague but he was trying to do the analysis at gene level not transcript level. Eventually I found those RSEM files…

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A natural product YSK-A blocks SARS-CoV-2 propagation by targeting multiple host genes

Hoareau, L. & DaSilva, E. J. Medicinal plants: A re-emerging health aid. Electron. J. Biotechnol. 2, 3–4 (1999). Google Scholar  Jahan, I. & Ahmet, O. Potentials of plant-based substance to inhabit and probable cure for the COVID-19. Turk. J. Biol. 44, 228 (2020). Article  CAS  PubMed  PubMed Central  Google Scholar …

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An issue with gtf file (ballgownrsem)

An issue with gtf file (ballgownrsem) 0 Hi everyone, When I tried to run ballgownrsem I encountered an issue which was caused by the inappropriately structured GTF file. Also, I tried to run code which is part of ballgownrsem and did not find the root of the issue. GTF files…

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Can I use TCGA-LUAD RNAseq count that had already normalized by RSEM in Limma-voom

Can I use TCGA-LUAD RNAseq count that had already normalized by RSEM in Limma-voom 0 Hi everyone, first of all, I’m new for bioinformatics. I have downloaded RNAseq data of TCGA-LUAD from UCSC that had already normalized RSEM normalized count and log2 transformed (log2 normcount+1). i wonder if i can…

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Chromosome-scale genome of the human blood fluke Schistosoma mekongi and its implications for public health | Infectious Diseases of Poverty

Barnett R. Schistosomiasis. (1474–547X (Electronic)). Steinmann P, Keiser J, Bos R, Tanner M, Utzinger J. Schistosomiasis and water resources development: systematic review, meta-analysis, and estimates of people at risk. Lancet Infect Dis. 2006;6(7):411–25. Article  PubMed  Google Scholar  Uthailak N, Adisakwattana P, Thiangtrongjit T, Limpanont Y, Chusongsang P, Chusongsang Y, et…

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using RSEM with non Trinity assembly

using RSEM with non Trinity assembly 0 Hi all, I am trying to use RSEM to receive relative abundance estimates of viruses within my metagenomic data, not transcripts. The problem is that I am no longer using Trinity as the assembler because I had much better luck with SPAdes, especially…

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Help doing differential expression analysis -experimental design and gProfiler TF interpretation-

Help doing differential expression analysis -experimental design and gProfiler TF interpretation- 0 Hi! I’m trying to do a differential expression analysis using breast cancer TCGA data. Firstly, I split the breast cancer cohort into two groups based on the expression level in z-score of a particular gene I’m interested in,…

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Functional convergence of genomic and transcriptomic architecture underlies schooling behaviour in a live-bearing fish

Krause, J. & Ruxton, G. D. Living in Groups (Oxford Univ. Press, 2002). Réale, D., Reader, S. M., Sol, D., McDougall, P. T. & Dingemanse, N. J. Integrating animal temperament within ecology and evolution. Biol. Rev. 82, 291–318 (2007). Article  PubMed  Google Scholar  Gartland, L. A., Firth, J. A., Laskowski,…

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coupling Cufflinks results with RSEM

coupling Cufflinks results with RSEM 0 Hello! I am opening this post to ask if I can use the transcript-level assembly obtained with Cufflinks (using a reference genome) to independently quantify the abundance levels of these sequences with RSEM (transcript-level quantification), specifically designed to quantify isoforms. Subsequently, I would perform…

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Biological and genetic characterization of a newly established human external auditory canal carcinoma cell line, SCEACono2

Ethic statement The Clinical Research Ethics Review Committee of Kyushu University Hospital approved the study (permit no. 29-43, 30-268, and 700-00). Written informed consent for the current research project was obtained before the tumor tissue, and a blood sample were harvested. This study was also conducted according to the principles…

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Issues while running abundance_estimates_to_matrix.pl

Issues while running abundance_estimates_to_matrix.pl 0 Hello,I am trying to generate a count matrix using abundance_estimates_to_matrix.pl script in Trinity, but I keep running into the same error. /usr/lib/trinityrnaseq/util/abundance_estimates_to_matrix.pl –est_method RSEM –gene_trans_map –name_sample_by_basedir GSNO_rep1/RSEM.isoforms.results GSNO_rep2/RSEM.isoforms.results GSNO_rep3/RSEM.isoforms.results wt_rep1/RSEM.isoforms.results wt_rep2/RSEM.isoforms.results wt_rep3/RSEM.isoforms.results -reading file: GSNO_rep1/RSEM.isoforms.results -reading file: GSNO_rep2/RSEM.isoforms.results -reading file: GSNO_rep3/RSEM.isoforms.results -reading file: wt_rep1/RSEM.isoforms.results -reading…

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Whole-genome sequencing analysis of suicide deaths integrating brain-regulatory eQTLs data to identify risk loci and genes

Li QS, Shabalin AA, DiBlasi E, Gopal S, Canuso CM, FinnGen ISGC, et al. Genome-wide association study meta-analysis of suicide death and suicidal behavior. Mol. Psychiatry 2023;28:891–900. McGuffin P, Marusic A, Farmer A. What can psychiatric genetics offer suicidology? Crisis. 2001;22:61–65. Article  CAS  PubMed  Google Scholar  Pedersen NL, Fiske A….

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TPM from STAR output without re-allign the file using RSEM or Salmon

Hi, I want to get the TPM files from aligned files generate with STAR and reading I found out that the easiest way is using RSEM or Salmon. My code for the alignment is /Users/c/STAR/bin/MacOSX_x86_64/STAR runThreadN 4 –genomeDir /Users/c/Desktop/Human_genome_index –readFilesIn /Users/c/Desktop/test/C1D20_R1_001_paired.fastq /Users/c/Desktop/test/C1D20_R2_001_paired.fastq –quantMode TranscriptomeSAM GeneCounts –outFileNamePrefix C1D20 –outSAMtype BAM SortedByCoordinate…

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RSEM not giving .genes.results and isoforms.results ; Plase check if you provide correct parameters/options for the pipeline!

RSEM not giving .genes.results and isoforms.results ; Plase check if you provide correct parameters/options for the pipeline! 0 I’m running RSEM for obtaining the final files of genes, isoforms and logs, etc. But it throws error as below after processing all rsem steps at rsem-run-em stage. Warning : The alignment…

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TruSeq strand-specificity in rsem-calculate-expression

TruSeq strand-specificity in rsem-calculate-expression 3 Hello, I’m trying to figure out the settings for analysing strand-specific RNA-Seq data from Illumina TruSeq chemistry with rsem-calculate-expression Specifically, regarding the –forward-prob parameter, the manual has this to say: ….Set to 1 for a strand-specific protocol where all (upstream) reads are derived from the…

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Salmon alignment based quantification

Salmon alignment based quantification 1 Hi, I aligned my sequences with STAR alignment and I need to quantify them with salmon with alignment based quantification mode. So I wonder > ./bin/salmon quant -t transcripts.fa -l <LIBTYPE> -a aln.bam -o salmon_quant In the command above, what should I write to transcripts.fa…

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How to check RNAseq support for annotated genes?

How to check RNAseq support for annotated genes? 2 Hello All, I have a set of annotated genes in gff3 format and corresponding RNA-seq data. What is the recommended approach and are there specific tools and parameters to determine the percentage of genes supported by the RNA-seq data?” Regards, B…

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Ballgown Error

Hi. I am using ballgown to support my reading analysis with Cufflinks as well as set up phenotype data for RSEM later. I was looking it up on Google and nobody seemed to have an answer with most of the questions being ignored. The error comes from this: bg_HNSCC =…

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What gene annotation was used for PanCanAtlas EBPlusPlus-corrected RNA-seq TCGA dataset?

What gene annotation was used for PanCanAtlas EBPlusPlus-corrected RNA-seq TCGA dataset? 0 Is there any info about what gene annotation (eg. genecode v26, ensembl 75, etc.) has been used to generate the PanCanAtlas EBpluPlus-corrected RNA-seq TCGA dataset (that is ebplusplusadjustpancan_illuminahiseq_rnaseqv2.geneexp.tsv from www.synapse.org/#!Synapse:syn4976363 )? RNAseq RSEM TCGA • 16 views Read…

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Iso-Seq method outperforms other long-read methods in benchmarking consortium study

A benchmarking of long-read RNA sequencing methods and analysis tools The Long-read RNA-Seq Genome Annotation Assessment Project (LRGASP) consortium, an initiative to systematically evaluate methods for transcript identification and quantification, recently released their final assessment of long-read sequencing technologies and tools in the preprint “Systematic assessment of long-read RNA-seq methods…

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Bioconductor Biovi

Comment: Handle zero effective gene length when tximport RSEM results by swbarnes2 &starf; 1.3k Are you sure that those genes with size < 1 have non-zero counts? Comment: P-value inflation? by James W. MacDonald 63k The table indicates that you have – 9122 genes with p>0.05 and FDR >0.1 –…

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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…

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Unsupervised clustering on gene expression data

Clustering is a data mining method to identify unknown possible groups of items solely based on intrinsic features and no external variables. Basically, clustering includes four steps: 1) Data preparation and Feature selection, 2) Dissimilarity matrix calculation, 3) applying clustering algorithms, 4) Assessing cluster assignment I use an RNA-seq dataset…

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Comparative transcriptome profiling to unravel the key molecular signalling pathways and drought adaptive plasticity in shoot borne root system of sugarcane

Shrivastava, A. K. & Srivastava, S. Sugarcane: Physiological and molecular approaches for improving abiotic stress tolerance and sustaining crop productivity. (ed. Narendra Tuteja et al.) 885–992 (Wiley Blackell, Weinheim, 2012). Reyes, J. A. O., Casas, D. E., Gandia, J. L. & Delfin, E. F. Agricultural Research Updates Vol. 35 (ed…

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Vertical and horizontal gene transfer shaped plant colonization and biomass degradation in the fungal genus Armillaria

Baumgartner, K., Coetzee, M. P. A. & Hoffmeister, D. Secrets of the subterranean pathosystem of Armillaria: subterranean pathosystem of Armillaria. Mol. Plant Pathol. 12, 515–534 (2011). Article  PubMed  PubMed Central  Google Scholar  Heinzelmann, R. et al. Latest advances and future perspectives in Armillaria research. Can. J. Plant. Pathol. 41, 1–23…

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RSEM uses different gene lengths for each sample.

RSEM uses different gene lengths for each sample. 0 I have 30 samples worth of unstranded RNA-seq data. I mapped with STAR on each of these samples and quantified with rsem-calculate-expression in RSEM based on the resulting bam files. The **.genes.results file for each sample showed that the same gene_id…

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Solution for pop-up problem with rsem-prepare-reference (RSEM can not recognize reference sequence name ATM…)

Solution for pop-up problem with rsem-prepare-reference (RSEM can not recognize reference sequence name ATM…) 0 Hello everyone, I was trying to use rsem-calculate-expression with the Aligned.toTranscriptome.out.bam file created by STAR. However, when I tried to run the code, I faced the error saying “RSEM can not recognize reference sequence name…

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Revisit where to find CCLE RNAseq in FPKM or RPKM using RSEM values to perform normalization- as was never answered usefully

Revisit where to find CCLE RNAseq in FPKM or RPKM using RSEM values to perform normalization- as was never answered usefully 0 I would like to find the CCLE RNA expression file that has either effective gene sizes or FPKM /RPKM (where estimated RSEM values have been used) to do…

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Deseq2 questions

Deseq2 questions 0 Hello everyone, I would like to perform a differential gene analysis on my dataset. It is composed of 8 pH points 5 different bacterial cocultures 3 biological triplicates = 120 samples. I have performed a de novo metatranscriptome with rnaSPAdes and now I am preparing my data…

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Is RSEM required for RNA-seq data analysis using STAR and edgeR?

Is RSEM required for RNA-seq data analysis using STAR and edgeR? 1 I have fastq files (n=40) obtained by paired-end unstranded RNA-seq. I would like to analyze these files using STAR for mapping and perform the differential expression analysis between the two groups by edgeR. I plan to apply TPM…

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Coevolution of the Tlx homeobox gene with medusa development (Cnidaria: Medusozoa)

Chang, E. S. et al. Genomic insights into the evolutionary origin of Myxozoa within Cnidaria. Proc. Natl Acad. Sci. USA 112, 14912–14917 (2015). Article  CAS  PubMed  PubMed Central  Google Scholar  Boero, F. et al. Inconsistent Evolution and Paedomorphosis among the Hydroids and Medusae of the Athecatae/Anthomedusae and the Thecatae/Leptomedusae (Cnidaria,…

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Importing RSEM processed data already formatted as a summarized experiment into DESeq2

Hello, I have a fairly simple question that I know has been addressed many times: I want to import RSEM data into DESeq2 for modeling and DE. For reproducibility, the workflow is: Unfortunately, it is costing me an inordinate amount of time, and I cannot find a perfectly analogous example…

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PCA for QC should include subset of top variable genes or full set of genes in RNAseq data?

PCA for QC in RNA seq data is to detect sample level outliers and batch effects etc. I am well aware that plotPCA of DESeq2 uses topn genes for doing PCA. My naive question from whole community is that do we need to use all genes or top highly variable…

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B-cell-specific checkpoint molecules that regulate anti-tumour immunity

Topalian, S. L. et al. Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. N. Engl. J. Med. 366, 2443–2454 (2012). Article  CAS  PubMed  PubMed Central  Google Scholar  Wolchok, J. D. et al. Nivolumab plus ipilimumab in advanced melanoma. N. Engl. J. Med. 369, 122–133 (2013). Article  CAS  PubMed …

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Y chromosome loss in cancer drives growth by evasion of adaptive immunity

Caceres, A., Jene, A., Esko, T., Perez-Jurado, L. A. & Gonzalez, J. R. Extreme downregulation of chromosome Y and cancer risk in men. J. Natl Cancer Inst. 112, 913–920 (2020). Article  PubMed  PubMed Central  Google Scholar  Kido, T. & Lau, Y. F. Roles of the Y chromosome genes in human…

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Pre-filtering of annotated isoforms based on splice junctions

Pre-filtering of annotated isoforms based on splice junctions 0 Is there a method to exclude transcript isoforms without any splicing junction support from my RNA-seq data? I would like to perform some pre-filtering of annotated isoforms before using Kallisto or RSEM. Thanks! isoforms RSEM splicing RNA-seq Kallisto • 15 views…

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RSEM implementation

RSEM implementation 0 I have the virus genome(fasta) and gff file and I am trying to prepare-reference through the following commands: rsem-prepare-reference –gff3 KT992094.1.gff3 KT992094.1.fasta or rsem-prepare-reference –gff3 KT992094.1.gff \ –gff3-genes-as-transcripts \ –bowtie \ KT992094.1.fasta \ ref/virus But it’s saying: Invalid number of arguments! How can I solve this issue?…

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Options to use with inward stranded reverse (ISR) reads

Options to use with inward stranded reverse (ISR) reads 0 Hi, I would like to perform a differential gene expression analysis on my dataset. The dataset is made up of paired-end files, and the libraries are ISR type (inward stranded reverse reads). I’d like to be sure that I’m including…

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Bioinformatics Approaches for Next-Generation Sequencing Analysis

Unlocking the Secrets of Genomic Data: Advanced Bioinformatics Techniques for Next-Generation Sequencing Analysis Navigating the Data Deluge: Bioinformatics Approaches for Next-Generation Sequencing Analysis Unlocking the Secrets of Genomic Data: Advanced Bioinformatics Techniques for Next-Generation Sequencing Analysis The advent of next-generation sequencing (NGS) technologies has revolutionized the field of genomics,…

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Chromatin reprogramming and bone regeneration in vitro and in vivo via the microtopography-induced constriction of cell nuclei

Dahl, K. N., Ribeiro, A. J. S. & Lammerding, J. Nuclear shape, mechanics, and mechanotransduction. Circ. Res. 102, 1307–1318 (2008). Article  CAS  PubMed  PubMed Central  Google Scholar  Alisafaei, F., Jokhun, D. S., Shivashankar, G. V. & Shenoy, V. B. Regulation of nuclear architecture, mechanics, and nucleocytoplasmic shuttling of epigenetic factors…

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Obtain a –gene-trans-map file after rnaSPAdes

Obtain a –gene-trans-map file after rnaSPAdes 0 Hello, I would like to perform a differential gene analysis on a dataset of 120 paird-end files. I obtained a good de novo quality metatranscriptome using rnaSPAdes (83% of the reads aligned exactly 1 time with bowtie2). I would now like to quantify…

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LinkedOmics :: Data Download

RNAseq (HiSeq, Gene level, Tumor) Download RNAseq data RSEM upper-quartile normalized (Illumina HiSeq platform, Gene-level) gene Expression (RSEM-UQ, Log2(Val+1)) 140 28057 cct RNAseq (HiSeq, Gene level, Normal) Download RNAseq data RSEM upper-quartile normalized (Illumina HiSeq platform, Gene-level) gene Expression (RSEM-UQ, Log2(Val+1)) 21 28057 cct RNAseq (HiSeq, Gene level, Duct) Download…

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Alignment of case vs. control from different origin

Alignment of case vs. control from different origin 0 Hi Biostars! I am coming to you with a relatively simple question, but one that i have surprisingly not found and answer to. I am working with a case-cohort of samples that were prepared for RNA-seq as paired reads with a…

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RNAseq Data and Pipeline

RNAseq Data and Pipeline 1 Hello, I am currently trying to do differential gene expression with TCGA data imported with R (GDCquery). I want to utilize DESeq2 as it is widely used and want publication quality results. I am struggling to understand where “raw” counts are found for star –…

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Counting intronic reads in bulk RNA-seq

Counting intronic reads in bulk RNA-seq 0 My experience with single-cell RNA-seq shows that the inclusion of intronic reads improves the sensitivity for several genes of interest, which otherwise have zero expression when only exonic reads are considered. While single-cell sequencing quantifiers now often have options to count intronic reads,…

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How does salmon uses polyester?

How does salmon uses polyester? 1 Hi Salmon users or developers! In salmon paper, to evaluate the ground truth, it uses RSEM to a certain data and uses polyester to the output of RSEM. Why use polyester on the output of RSEM? In salmon github repo, can somebody direct me…

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Genecount-difference between HT-seq count, RSEM, and Kallisto

Genecount-difference between HT-seq count, RSEM, and Kallisto 0 Hi I ran three genecount software tools (ht-seq, RSEM, Kallisto) to calculate genecount of RNA-seq data. For Ht-seq, i used STAR aligned Transcriptomesortedcordinate.bam file and defautl MAPQ score with intersection_nonempty mode. For RSEM, i used STAR aligner (used .gtf for building reference)…

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Issue running RSEM

Issue running RSEM 0 I am trying to run RSEM on my output from STAR. I have vreated the RSEM ref and have it in my working diretory, however when I run RSEM: rsem-calculate-expression –paired-end –bam -p 16 “$f” RSEMref/ rsem_out/”$base” I get this error: rsem-parse-alignments RSEMref/ rsem_out/MR13_S13.temp/MR13_S13 rsem_out/MR13_S13.stat/MR13_S13 MR13_S13_mappedAligned.toTranscriptome.out.bam…

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Read counting in RSEM and Salmon (alignment mode)

Read counting in RSEM and Salmon (alignment mode) 1 Hi all, I was wondering if someone can clarify an issue for me. When counting reads for RNA-seq, you can use EM-based algorithms to correctly assign multimapping reads. Thus, if you have 5 single-end reads that map to 1 transcript, and…

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PRR15 deficiency facilitates malignant progression by mediating PI3K/Akt signaling and predicts clinical prognosis in triple-negative rather than non-triple-negative breast cancer

Cell culture and reagents Non-cancerous mammary epithelium cell (MCF10A), breast cancer cells including luminal (MCF7, MDA-MB-361, T47D, and BT474), HER2amp (SKBR3), and triple-negative (MDA-MB-231, CAL51, BT20, and MDA-MB-468) subtypes, as well as human embryonic kidney 293T (HEK-293T) cells, were purchased from the American Type Culture Collection (Manassas, VA, USA). MDA-MB-231,…

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Dryad | Data — RNAseq transcriptome of draining lymph node (LN) and tumor of MC38 murine tumors treated with cryoablation and chitosan/IL-12

Focal ablation technologies are routinely used in the clinical management of inoperable solid tumors but often result in incomplete ablations leading to high recurrence rates. Adjuvant therapies capable of safely eliminating residual tumor cells are therefore of great clinical interest. Interleukin 12 (IL-12) is a potent antitumor cytokine that can…

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RSEM expression values

RSEM expression values 0 In RSEM.isoform.TMM.EXPR.matrix, some values for a set of 3 samples are like this: 0.596 0.782 0 2.173 0 0 4465.372 3953.622 5033.097 Why there is so much difference between samples and among different transcripts? RSem expression Quantification Trinity • 26 views Login before adding your answer….

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Nextflow rnaseq finishing early

Nextflow rnaseq finishing early 0 Hi I’m running the RNA-seq pipeline from nextflow and I have been running it without problems until this dataset it just stops prematurely saying it has finished when it doesn’t even aligns the reads with salmon. Any ideas what may be going on? I have…

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Can I run DESeq2 with SMART-Seq data

Can I run DESeq2 with SMART-Seq data 1 @assa-yeroslaviz-1597 Last seen 2 days ago Germany my data set comes from single-cell SMART-Seq data, where I have in total 48 samples, 24 samples for the control and 24 for my KO. Each sample represent one cell. I have used STAR to…

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Add HI:i: tag to a BAM file

Add HI:i:<n> tag to a BAM file 0 Hi all, I’ve been using STAR in conjunction with RSEM to get the most accurate quantification of RNA-seq for a while now. However, in one of the recent projects, I needed to map reads to a repetitive reference, generating an alignment with…

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Converting an output de-novo transcriptome assembled with Trinity to a .gff3 file

Converting an output de-novo transcriptome assembled with Trinity to a .gff3 file 2 Hello! I’ve de-novo assembled a transcriptome from Trinity, resulting into Trinity.fasta, whose headers look like this: >TRINITY_DN29256_c0_g1_i1 len=323 path=[0:0-322] Followed, in the next line, by the sequence. To run an external downstream analysis with a R script,…

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Empty genes.bam files in RSEM-STAR workflow

Empty genes.bam files in RSEM-STAR workflow 0 This question concerns an RNAseq data aligment and transcript quantification step that generates empty bam files but still generates counts file. I would love to know 1. If the count data is reliable if bam files are empty ? and 2. How can…

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Answer: using Firebrowser to identify disease type

The solution to this is within the `Samples.mRNASeq` that gives data which can be saved in JSON format: [0] { cohort “ACC”, expression_log2 3.635731, gene “CD274”, geneID 29126, protocol “RSEM”, sample_type “TP”, tcga_participant_barcode “TCGA-PK-A5HB”, z-score -0.01802174 }, [1] { cohort “ACC”, expression_log2 2.725785, gene “CD274”, geneID 29126, protocol “RSEM”, sample_type…

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Cross-platform normalization enables machine learning model training on microarray and RNA-seq data simultaneously

We aimed to assess the extent to which it was possible to effectively normalize and combine microarray and RNA-seq data with existing methods for use as a training set for machine learning applications. We assessed performance on holdout sets composed entirely of microarray data and entirely of RNA-seq data. To…

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Information on “sample_name.cnt” obtained by an RSEM analysis

Information on “sample_name.cnt” obtained by an RSEM analysis 1 Hello, I obtained a “sample_name.cnt” in a newly created “sample_name.stat” directory after an RSEM-1.3.3 analysis. Shown below is the content of the “sample_name.cnt”. What do these numbers mean? Thank you in advance for your kindness. 0 2726098 0 2726098 1534055 1192043…

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RSEM calculate-expression has only one sample in .isoforms.results file

RSEM calculate-expression has only one sample in .isoforms.results file 1 Hello, I think I have a few misunderstandings about how to use RSEM, and have provided my script below. I have three questions: My script outputted a .isoform.results file, but with only one sample in it. As you can see…

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Question about RNA-Seq data alignment

Question about RNA-Seq data alignment 3 Hi, I have a question about genome alignment. I am working with RNA-Seq dataset to study the impact of Liquid Culture in response to virus of different doses in Human. I was exploring what could be the good strategy or best in practice method…

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Matching IDs between 3+ files and specifying output using dictionaries in Python

Hello all, I have a code that is supposed to read a file ‘filecontig,’ take all the sequence IDs within that file, match those IDs to IDs in files ‘filetaxa’ and ‘fileTPM’ and output the taxonomical classifications as well as the transcripts per million that match each respective ID. I…

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mapping – STAR error in snakemake pipeline: “EXITING because of FATAL ERROR: could not open genome file”

I’m trying to use a 2 pass STAR mapping strategy (also explained here informatics.fas.harvard.edu/rsem-example-on-odyssey.html), but I’m getting an error. I’ve read through this page [https://github.com/alexdobin/STAR/issues/181] and I have a similar issue, but the discussed solutions don’t seem to help. Perhaps this is more a snakemake issue rather than a STAR…

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In vivo transomic analyses of glucose-responsive metabolism in skeletal muscle reveal core differences between the healthy and obese states

Animals and sample preparation Animal experiments were performed as previously described12. C57BL/6J WT mice or ob/ob mice at ten weeks of age were purchased from Japan SLC Inc. (Shizuoka, Japan). The phenotypic data of the mice are summarized in Table S1. Animal experiments were approved by the animal ethics committee…

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Finding DEGs from HISAT2/STRINGTIE output

Finding DEGs from HISAT2/STRINGTIE output 0 Hello, I have to search for DEGs from four samples of crop. I am following reference based mapping of reads to genome using HISAT2. I have completed till the generation of merged .gtf files for the samples using STRINGTIE. Since I am new to…

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Use RSEM and Bowtie2 to align paired-end sequences

Use RSEM and Bowtie2 to align paired-end sequences 0 I want to use rsem-calculate-expression and bowtie2 aligner to align paired-end sequence based on the following conditions: 2 processors generate BAM file very fast bowtie2 sensitivity append gene/transcript name My code: rsem-refseq-extract-primary-assembly GCF_000001405.31_GRCh38.p5_genomic.fna GCF_000001405.31_GRCh38.p5_genomic.primary_assembly.fna rsem-prepare-reference –gff3 GCF_000001405.31_GRCh38.p5_genomic.gff –bowtie2 –bowtie2-path /bowtie2-2.4.5-py39hd2f7db1_2 –trusted-sources…

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Bioinformatics analysis identifies widely expressed genes

1Department of Orthopedics, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, People’s Republic of China; 2Department of Pediatrics, The Shanxi Medical University, Taiyuan, Shanxi, People’s Republic of China Correspondence: Jun Qian, Department of Orthopedics, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui,…

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how to build index for cdna?

Hello, I can build index for Mus_musculus.GRCm38.dna_sm.toplevel.fa, but when build for Mus_musculus.GRCm38.cdna.all.fa, there is a bug: “rsem-extract-reference-transcripts Mus_musculus.GRCm38.cdna.all.fa 0 Mus_musculus.GRCm38.cdna.all.fa.gtf None 0 Mus_musculus.GRCm38.cdna.all” failed! Plase check if you provide correct parameters/options for the pipeline! Traceback (most recent call last): File “../indrops.py”, line 1770, in project.build_transcriptome(args.genome_fasta_gz, args.ensembl_gtf_gz, mode=args.mode) File “../indrops.py”, line…

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RNAseq data DEG analysis – DESeq2 normalized data

RNAseq data DEG analysis – DESeq2 normalized data 1 1) You can’t use because those data are already normalized and log-transformed. 3) RSEM expected_count is best to start off with for differential expression. Login before adding your answer. Traffic: 2089 users visited in the last hour Read more here: Source…

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3 -tag XM” failed! when running rsem-calculate-expression

Dear sir, When I ran “rsem-calculate-expression –paired-end –alignments -p 8input.bam” gencodev22 ./out. I got error message rsem-parse-alignments ../bowtie2/hg38 ./rsem-out.temp/rsem-out ./rsem-out.stat/rsem-out /NGS_Storage/Debbie/RNA-seq/variant_calling_20210602/RNA-leukemia002A-906.para.bam 3 -tag XM Read A00355:209:H3KTLDSX2:2:2606:24677:17425: The adjacent two lines do not represent the two mates of a paired-end read! (RSEM assumes the two mates of a paired-end read should…

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rsem-calculate-expression error /lib/libc.so.6: version `GLIBC_2.14′ not found

Hi, When I run rsem-calculate-expression. I got an error like this: $ home/trinityrnaseq/trinity-plugins/rsem/rsem-parse-alignments /home/trinityrnaseq/trinity-plugins/rsem/rsem-parse-alignments: /lib/libc.so.6: version GLIBC_2.15′ not found (required by /home/trinityrnaseq/trinity-plugins/rsem/rsem-parse-alignments) /home/trinityrnaseq/trinity-plugins/rsem/rsem-parse-alignments: /lib/libc.so.6: version GLIBC_2.14′ not found (required by /home/trinityrnaseq/trinity-plugins/rsem/rsem-parse-alignments) Any help would be greatly appreciated~~ Read more here: Source link

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How to reduce the impact of one varaible in Deseq2 or edgeR for multivariate value analysis?

Hello, everyone, I’m recently meeting this problem with my analysis, which i’ve done a lots of research and asked people around but their answers are quite confusing, so if I can get more opinions, that’d be terrific and thanks at advance. So I’m doing an analysis of DEGs using Deseq2…

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CIBERSORTxFractions ERROR: Could not read /src/outdir//temp.Fractions.simfracs.tsv

Hello, can anyone offer any insight into the following problem? I am trying to run the following CIBERSORTx function locally: docker run -v /media/mark/seagate2/data/CIBERSORTx_GC:/src/data -v /media/mark/seagate2/data/CIBERSORTx_GC:/src/outdir cibersortx/fractions –username <my_user_name> –token <my_token> –single_cell TRUE –refsample reference.txt –mixture rsem_mixture_TPM.tsv –fraction 0 –rmbatchSmode TRUE I get the following output to the terminal: >Running…

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DESeq2 input from GDAC firehose

Hi guys, I hope you are fine. I’m not good in English so if you couldn’t understand my question, please feel free to reply. I’m a beginner of bioinformatics. I want to practice differential expressed gene (DEG) analysis in R. The RNA seq data I used was downloaded from broad…

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Comparative de novo transcriptome analysis identifies salinity stress responsive genes and metabolic pathways in sugarcane and its wild relative Erianthus arundinaceus [Retzius] Jeswiet

1. Singh, A. et al. Phytochemical profile of sugarcane and its potential health aspects. Pharmacogn. Rev. 9, 45–54 (2015). CAS  PubMed  PubMed Central  Google Scholar  2. Eggleston, G. Positive aspects of cane sugar and sugar cane derived products in food and nutrition. J. Agric. Food Chem. 66, 4007–4012 (2018). CAS …

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rsem-calculate-expression with STAR takes forever but without error

rsem-calculate-expression with STAR takes forever but without error 1 Hi all, I am running rsem-calculate-expression after succesfully generating the reference files with rsem-prepare-reference. However, the command never proceeds past: “started mapping” like in the example below Sep 16 08:44:43 ….. started STAR run Sep 16 08:44:43 ….. loading genome Sep…

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STAR+RSEM pippline without gtf

STAR+RSEM pippline without gtf 0 Dear all, I have question I mapped reads on cds sequence through STAR I don’t have gtf file and want to calculate read count using RSEM but I am stuck by error “RSEM error: RSEM currently does not support gapped alignments” as I don’t have…

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Download TCGA and GTEX data from Xena toilHub for (full genome but for 1 cancer/tissue type)

Download TCGA and GTEX data from Xena toilHub for (full genome but for 1 cancer/tissue type) 0 Dear All, I would like to download TCGA and GTEX gene expression data for ovarian cancer and ovary respectively from the Xena toilHub platform (all genes; RSEM expected counts). However, I only found…

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Survival analysis for a list of genes using TCGA data

Survival analysis for a list of genes using TCGA data 1 Hi all, I have a list of genes (around 300 genes) and I want survival analysis to find only significant genes. I am using TCGA RSEM normalized data in survival package using following command, but I’m not sure how…

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pughlab/inspire-genomics: Pan-cancer analysis of genomic and immune landscape profiles of metastatic solid tumors treated with pembrolizumab

Contents Serial circulating tumor DNA (ctDNA) monitoring is emerging as a non-invasive strategy to predict and monitor immune checkpoint blockade (ICB) therapeutic efficacy across cancer types. Yet, limited data exist to show the relationship between ctDNA dynamics and tumor genome and immune microenvironment in patients receiving ICB. Here, we present…

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regarding coexpression analysis (by pearson method) of genes shortlisted from expression profile of a dataset obtained from GDAC Firehouse database

regarding coexpression analysis (by pearson method) of genes shortlisted from expression profile of a dataset obtained from GDAC Firehouse database 0 Hello guys, Should i consider genes having “zero” normalized count given in expression data of a dataset while doing coexpression analysis with respect to a particular gene (whose expression…

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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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rsem-prepare-reference –allele-to-gene-map (file information required)

rsem-prepare-reference –allele-to-gene-map <file> (file information required) 0 –allele-to-gene-map <file>Use information from <file> to provide gene_id and transcript_id information for each allele-specific transcript. Each line of <file> should be of the form: gene_id transcript_id allele_id with the fields separated by a tab character. This option is designed for quantifying allele-specific expression….

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