Seeding nucleofected hPSCs in 96-well plates using limited dilution
By savannah onThis protocol describes a standard procedure for seeding nucleofected human pluripotent stem cells (hPSCs) in 96-well plates using limited dilution. This protocol follows nucleofection of hPSCs as described in detail in the collection “Nucleofection (Amaxa) and electroporation (Biorad) of hPSCs.” dx.doi.org/10.17504/protocols.io.b4qnqvve
Sample preparation for aCGH karyotyping
By savannah onThis protocol describes the standard procedure preparing cell pellets from human pluripotent stem cells (hPSCs) cultured on MEFs for outsourced aCGH karyotyping.
Preparing samples for NGS
By savannah onThis protocol describes a standard procedure used to prepare PCR samples for Next Generation Sequencing (NGS).
Preparing plasmids for nucleofection of hPSCs
By savannah onThis protocol describes the standard procedure for the preparation of plasmids to be delivered into human pluripotent stem cells (hPSCs) using nucleofection.
Preparing of genomic DNA from in vitro cultured cells
By savannah onThis protocol describes a standard procedure for preparing crude cell lysate which can be further analyzed by PCR.
Preparing mRNA for nucleofection of hPSCs
By savannah onThis protocol describes the standard procedure for preparing mRNA to be delivered into human pluripotent stem cells (hPSCs) using nucleofection.
Preparing MEF-cultured hPSCs for nucleofection
By savannah onThis protocol describes the procedure or preparing MEF-cultured human pluripotent stem cells (hPSCs) for the delivery of plasmids, mRNA, or ribonucleoprotein (RNP) using nucleofection.
Preparing feeder-free hPSCs for nucleofection
By savannah onThis protocol describes the standard procedure preparing feeder-free human pluripotent stem cells (hPSCs) for nucleofection.
Preparation of mouse embryonic fibroblast (MEF) feeder plates for hPSC cultures
By savannah onThis protocol describes the preparation of mouse embryonic fibroblasts (MEFs) as feeder cells for human pluripotent stem cell (hPSC) culture.
Preparation of LRRK2 RCKW cryo-EM grids
By savannah onThis is Leschziner's Lab updated protocol for making cryo-EM grids for LRRK2 RCKW. This protocol, when using lower protein concentration, results in better monomer and dimer formation than the old protocol.
Pluripotency markers staining
By savannah onThis protocol describes the standard procedure for staining pluripotency markers, e.g., OCT4, SSEA4, alkaline phosphatase, and etc. on human pluripotent stem cells (hPSCs).
Passaging of feeder-free hPSCs
By savannah onThis protocol describes the process of passaging human pluripotent stem cells (hPSCs) for feeder-free culturing of hPSCs using Accutase or ReLeSF.
Nucleofection of hPSCs
By savannah onThis protocol describes the standard procedure for the delivery of plasmids, mRNA, or ribonucleoprotein (RNP) into human pluripotent stem cells (hPSCs) using nucleofection.
Nucleofection (Amaxa) and electroporation (Biorad) of hPSCs
By savannah onThis collection describes the standard procedure for the delivery of plasmids, mRNA, or ribonucleoprotein (RNP) into human pluripotent stem cells (hPSCs) using nucleofection or electroporation.
Mitomycin C inactivation of mouse embryonic fibroblasts (MEFs) for hPSC cultures
By savannah onThis protocol describes the process of using Mitomycin C to inactivate mouse embryonic fibroblasts (MEFs), which can then be used as feeder cells for human pluripotent stem cell (hPSC) culture.
Maintenance and inactivation of mouse embryonic fibroblasts (MEFs) as feeder cells for human pluripotent stem cell culture
By savannah onThis collection describes the maintenance and inactivation of mouse embryonic fibroblasts (MEFs) as feeder cells for human pluripotent stem cell (hPSC) culture.
Lentivirus production
By savannah onThis protocol is associated with the following preprint, published on February 19, 2022: “The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds.”
In vitro synthesis of PE2 (nCas9-MMLV RT fusion) polyA mRNA using T7 RNA polymerase
By savannah onWe describe the preparation of synthetic capped and polyadenylated messenger RNA (mRNA) encoding a Cas9 nickase-reverse transcriptase fusion protein (PE2) for use in genome editing with a method called prime editing.
In vitro assembling of RNP for nucleofection of hPSCs
By savannah onThis protocol describes the procedure for the in vitro assembly of ribonucleoprotein (RNP) which can be delivered into human pluripotent stem cells (hPSCs) using nucleofection.