Optimized single-cell RNA sequencing protocol to study early genome activation in mammalian preimplantation development

Abstract

Summary: Here, we present a modification of single-cell tagged reverse transcription protocol to study gene expression on a single-cell level or with limited RNA input. We describe different enzymes for reverse transcription and cDNA amplification, modified lysis buffer, and additional clean-up steps before cDNA amplification. We also detail an optimized single-cell RNA sequencing method for handpicked single cells, or tens to hundreds of cells, as input material to study mammalian preimplantation development.For complete details on the use and execution of this protocol, please refer to Ezer et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Bioinformatics
• Sequence analysis
• Single Cell
• Developmental biology
• Genomics
• Sequencing