Single cell transcriptomics – a gerneric explanation of common steps

Single-cell RNA sequencing (scRNA-seq) has significantly advanced our knowledge of biological systems by studying the single cell transcriptome.

Compared to bulk RNA-seq, scRNA-seq is able to study cellular heterogeneity. BD^TM Rhapsody

Steps of single cell transcriptomics

Capture single cells
Magnetic barcoded beads to microwells (one bead to one cell)
Cell lysis to hybridize mRNA´s onto barcoded beads by polyA-tail
Retrieve beads and start library preparation with reverse transcription
Amplifications and quality control of the library
Adding illumine sequencing primers
Illumina sequencing
Pre-processing of raw data to count matrics, quality control, normalization, data correction, feature selection
Downstream analsysis e.g. clustering, cluster annotation, trajectory inference, compositional analysis, differential expression