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Catalog
ASAP is committed to accelerating the pace of discovery and informing a path to a cure for Parkinson’s disease through collaboration, research-enabling resources, and data sharing. We’ve created this catalog to showcase the research outputs and tools developed by ASAP-funded programs.
Protocol
Standard operating procedure for the isolation of genetically engineered hPSCs clones in a high-throughput way
This collection describes a standard procedure for isolating single human pluripotent stem cell (hPSC) clones in a high-throughput way. This collection 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
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Amplifying the target genomic region by PCR
This protocol describes a procedure for amplifying targeted genomic regions using a PCR reaction.
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AAVS1 Knock-in
This protocol describes the standard procedure to knock-in constructs to the AAVS1 safe harbor locus in hPSCs.
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Adapting hPSCs cultured on MEFs to feeder-free system
This protocol describes the procedure of adapting human pluripotent stem cells (hPSCs) to feeder-free culturing conditions using mTeSR-plus or StemFlex.
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Purification of the PE2 nCas9-RT protein
This protocol describes the process of expressing and purifying the nicking Cas9-MMLV RT fusion protein for prime editing.
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CRISPResso analysis
This protocol describes the process of using CRISPResso to analyze Next Generation Sequencing (NGS) data from CRISPR-targeted, genome-edited samples.
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Culture and transfection of HEK293T cells
This protocol describes a standard procedure culturing and transfecting HEK293T cells. Protocol overview: A. Culturing HEK293T cells; B. Transfection of HEK293T cells with Lipofectamine 2000.
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Duplicating 96-well plate-cultured hPSCs clones
This protocol describes a standard procedure for duplicating 96-well plate-cultured human pluripotent stem cells (hPSCs).
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Electroporation of hPSCs
This protocol describes the standard procedure for the delivery of plasmids into human pluripotent stem cells (hPSCs) using electroporation.
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Expansion of mouse embryonic fibroblasts (MEFs) for hPSC cultures
This protocol describes the expansion of mouse embryonic fibroblasts (MEFs) as feeder cells for human pluripotent stem cell (hPSC) culture.
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FACS-based enrichment of transfected hPSCs
This protocol describes the procedure FACS-based enrichment of transfected human pluripotent stem cells (hPSCs).
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Feeder-free culturing of hPSCs
This collection describes the standard procedure of feeder-free culturing of human pluripotent stem cells (hPSCs) using mTeSR-plus or StemFlex.
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Freezing of feeder-free hPSCs
This protocol describes the process of freezing feeder-free human pluripotent stem cells (hPSCs) using Accutase or ReLeSR.
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Freezing of hPSCs grown on MEFs
This protocol describes the standard procedure of freezing human pluripotent stem cells (hPSCs), which were grown on inactivated mouse embryonic fibroblasts (MEFs).
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