−/−to obtain the 1× solution. Spheroids were then removed from the collagen gel if necessary and washed twice with 2 mM EDTA in PBS. Spheroids were then dissociated in 750 µl of 1× papain solution at 37 °C and 750 r.p.m. on a thermomixer . The enzymatic reaction was stopped with 750 µl of stop solution consisting of 1 mg ml.
The Cell Ranger pipeline was used to perform sample demultiplexing and barcode processing and to generate the single-cell gene counting matrix. Briefly, samples were demultiplexed to produce a pair of FASTQ files for each sample. Reads containing sequence information were aligned using the reference provided with Cell Ranger based on the GRCh37 reference genome and ENSEMBL gene annotation.
The count data matrix was read into R and used to construct a Seurat object . The Seurat package was used to produce diagnostic quality control plots and to select thresholds for further filtering. Filtering method was used to detect outliers and high numbers of mitochondrial transcripts. These preprocessed data were then analyzed to identify variable genes, which were used to perform a principal-component analysis .
dzhk_germany UMG_Tweet HelmholtzMunich Saar_Uni karolinskainst YaleSurgery Co-authors ab_meier and Alessandra Moretti go behindthepaper on ‘epicardioids’ that replicate the morphological and functional self-organization of ventricular myocardium and epicardium, opening new avenues for investigation
dzhk_germany UMG_Tweet HelmholtzMunich Saar_Uni karolinskainst YaleSurgery A mini-heart in a Petri dish NBTintheNews via TU_Muenchen
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