Single-Cell RNA-seq Technologies (10x Genomics, Smart-Seq, Drop-seq and more) | BioCode скачать в хорошем качестве

Single-Cell RNA-seq Technologies (10x Genomics, Smart-Seq, Drop-seq and more) | BioCode

In this video, we will dive into the world of Single-cell RNA sequencing (scRNA-seq), a powerful technique that enables researchers to study gene expression in individual cells. There are several technologies available for scRNA-seq, each with its own advantages and limitations. This video will focus on some of the most popular technologies used for scRNA-seq, including 10x Genomics, Smart-Seq, Drop-seq, Seq-Well, sci-RNA-seq, and CEL-Seq. 10x Genomics is a microfluidic system that partitions cells into droplets, with each droplet containing a bead with a unique barcode. The cells are then lysed, and the RNA is captured and sequenced using Illumina sequencing technology. 10x Genomics provides high-throughput and high-quality scRNA-seq data with relatively low input RNA requirements. Smart-Seq uses a template-switching mechanism to capture the full-length RNA transcripts from individual cells. The cells are lysed, and the RNA is captured and sequenced using Illumina sequencing technology. Smart-Seq provides high sensitivity and coverage, but requires more input RNA than other scRNA-seq methods. It is also more technically challenging and time-consuming. Drop-seq is a high-throughput scRNA-seq method that encapsulates individual cells into droplets along with barcoded beads, which capture the RNA transcripts from each cell. The resulting cDNA is then amplified and sequenced using Illumina sequencing technology. Drop-seq provides a high-throughput and cost-effective platform for scRNA-seq, with low input RNA requirements. Seq-Well is a high-throughput scRNA-seq method that uses a microfabricated array to capture individual cells in wells, along with barcoded oligonucleotides to capture RNA transcripts. The amplified cDNA is then sequenced using Illumina sequencing technology. Seq-Well provides high-throughput scRNA-seq data with low input RNA requirements. Other scRNA-seq technologies include sci-RNA-seq, which uses CRISPR-Cas9 technology to label RNA with a unique barcode in each cell, and CEL-Seq, which involves multiplexing cells and amplifying RNA using linear amplification and in vitro transcription. CEL-Seq provides high-throughput scRNA-seq data with low input RNA requirements, but has lower sensitivity and coverage than other scRNA-seq methods. Once scRNA-seq data has been generated, it needs to be analyzed using specialized bioinformatics tools. Common steps in scRNA-seq analysis include quality control and filtering of cells and transcripts, normalization of gene expression data, dimensionality reduction to visualize and cluster cells based on similarities in gene expression, and differential expression analysis to identify genes that are differentially expressed between cell types or conditions. There are many software tools available for scRNA-seq analysis, including Seurat, Cell Ranger, and STAR. In summary, scRNA-seq technologies have revolutionized our understanding of gene expression and have become an essential tool in many fields of biology. The ability to study gene expression at the single-cell level has opened up new avenues for research, enabling scientists to explore cellular heterogeneity and identify novel cell types and regulatory pathways.