GoT-Splice protocol for multi-omics profiling of gene expression, cell-surface proteins, mutational status, and RNA splicing in human cells
STAR PROTOCOLS(2024)
Abstract
Studying RNA splicing factor mutations is challenging due to difficulties in distinguishing wild -type and mutant cells within complex human tissues and inaccuracies associated with reconstructing splicing signals from short -read sequencing data. Here, we present Genotyping of Transcriptomes (GoT)-Splice, a protocol that overcomes these limitations by combining GoT with enhanced long -read single -cell transcriptome and cell -surface proteomics profiling. We describe steps for long -read library preparation and analysis, followed by cDNA re -amplification, enrichment of mutation of interest, sample indexing, and GoT library preparation. For complete details on the use and execution of this protocol, please refer to Corte s-Lo pez et al. 1
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Key words
Single Cell,Genomics,Sequencing,RNA-seq,Molecular Biology,Gene Expression
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