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. BDTM Rhapsody

Steps of single cell transcriptomics

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


Author: Kelly Blust (ESR11)