Nucleofection (Amaxa) and electroporation (Biorad) of hPSCs v2

This collection describes the standard procedure for the delivery of plasmids, mRNA or ribonucleoprotein (RNP) into human pluripotent stem cells (hPSCs) using nucleofection or electroporation. Collection Overview Preparing MEF-cultured hPSCs for nucleofection Preparing Feeder-free hPSCs for nucleofection Preparing plasmids for nucleofection Preparing mRNA for nucleofection In vitro assembling of RNP for nucleofection Nucleofection of hPSCs Electroporation of hPSCs A. Preparation of MEFs-cultured hPSCs for electroporation B. Preparing plasmids for electroporation C. Electroporation FACS-based enrichment of transfected cells A. Preparation of samples for FACS sorting B. After FACS General notes Throughout these protocols, the term hPSC is used to collectively refer to both hiPSCs and hESCs. All described procedures have been tested and work equally well for hiPSCs and hESCs.