Freezing of hPSCs grown on MEFs
By savannah onThis protocol describes the standard procedure of freezing human pluripotent stem cells (hPSCs), which were grown on inactivated mouse embryonic fibroblasts (MEFs).
Freezing of feeder-free hPSCs
By savannah onThis protocol describes the process of freezing feeder-free human pluripotent stem cells (hPSCs) using Accutase or ReLeSR.
Feeder-free culturing of hPSCs
By savannah onThis collection describes the standard procedure of feeder-free culturing of human pluripotent stem cells (hPSCs) using mTeSR-plus or StemFlex.
FACS-based enrichment of transfected hPSCs
By savannah onThis protocol describes the procedure FACS-based enrichment of transfected human pluripotent stem cells (hPSCs).
Expansion of mouse embryonic fibroblasts (MEFs) for hPSC cultures
By savannah onThis protocol describes the expansion of mouse embryonic fibroblasts (MEFs) as feeder cells for human pluripotent stem cell (hPSC) culture.
Electroporation of hPSCs
By savannah onThis protocol describes the standard procedure for the delivery of plasmids into human pluripotent stem cells (hPSCs) using electroporation.
Duplicating 96-well plate-cultured hPSCs clones
By savannah onThis protocol describes a standard procedure for duplicating 96-well plate-cultured human pluripotent stem cells (hPSCs).
Culture and transfection of HEK293T cells
By savannah onThis protocol describes a standard procedure culturing and transfecting HEK293T cells. Protocol overview: A. Culturing HEK293T cells; B. Transfection of HEK293T cells with Lipofectamine 2000.
Crystal structure of a substrate-trapping variant of PPM1H phosphatase
By savannah onCrystal structure of a substrate-trapping variant of PPM1H phosphatase (as reported in 10.15252/embr.202152675) deposited in the Protein Data Bank with code 7L4I.
Crystal structure ATG9 HDIR in complex with the ATG13:ATG101 HORMA dimer
By savannah onExperimental Data Snapshot Crystal structure ATG9 HDIR in complex with the ATG13:ATG101 HORMA dimer PDB DOI: 10.2210/pdb8DO8/pdb Classification: SIGNALING PROTEIN Organism(s): Homo sapiens Expression System: Homo sapiens Mutation(s): No Deposited: 2022-07-12 Released: 2022-11-23 Deposition Author(s): Buffalo, C.Z., Ren, X., Yokom, A.L., Hurley, J.H. Funding Organization(s): Other private Method: X-RAY DIFFRACTION Resolution: 2.41 Å R-Value Free: 0.251 R-Value Work: 0.204 R-Value Observed: 0.205
CRISPResso analysis
By savannah onThis protocol describes the process of using CRISPResso to analyze Next Generation Sequencing (NGS) data from CRISPR-targeted, genome-edited samples.
CRISPR/Cas9-based functional genomics in human induced pluripotent stem cell–derived models: Can “the stars align” for neurodegenerative diseases?
By savannah onViewpoint article on CRISPR/Cas9-based functional genomics in human induced pluripotent stem cell–derived models.
Coating plates
By savannah onThis protocol describes the process of coating plates using either VTN, Matrigell, or Geltrex for use in culturing of feeder-free human pluripotent stem cells (hPSCs).
Amplifying the target genomic region by PCR
By savannah onThis protocol describes a procedure for amplifying targeted genomic regions using a PCR reaction.
Adapting hPSCs cultured on MEFs to feeder-free system
By savannah onThis protocol describes the procedure of adapting human pluripotent stem cells (hPSCs) to feeder-free culturing conditions using mTeSR-plus or StemFlex.
AAVS1 Knock-in
By savannah onThis protocol describes the standard procedure to knock-in constructs to the AAVS1 safe harbor locus in hPSCs.
Single cell analysis of iPSC-derived midbrain organoids
By savannah onThe following script was used for analysis of gene corrected (GC) versus GBA1 mutant (MUT) midbrain organoids. The purpose was to combine, filter, integrate, and identify clusters and differentially expressed genes sets.
Western blotting to detect ATP13A2 and ATP13A3
By savannah onProtocol to detect ATP13A2 and ATP13A3 via Western blotting.
Microscopy-based mitochondrial morphology measurements in iNeurons
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Microscopy-based mtDNA turnover measurements in HeLa and iNeurons
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
