Recombinant:Article Title: Mapping Gene Circuits Essential for Germ Layer Differentiation via Loss-of-Function Screens in Haploid Human Embryonic Stem Cells.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies PerCP-Cy5.5-conjugated mouse anti-human SOX1 BD Biosciences Cat# 561549; RRID:AB_10694879 Alexa Flour 488-conjugated SOX17 BD Biosciences Cat# 562205; RRID:AB_10893402 goat anti-human/mouse NCAM1 R&D Systems Cat# AF2408; RRID: AB_442152 donkey anti-goat Cy5-conjugated secondary antibody Jackson ImmunoResearch Laboratories Cat# 705-175-147; RRID: AB_2340415 Chemicals, Peptides, and Recombinant Proteins mTeSR1 medium STEMCELL Technologies Cat# 05850 Y-27632-dihydrochloride Gentaur Cat# A3008 TrypLE Select Thermo Fisher Scientific Cat# 12563029 Knockout Dulbecco’s Modified Eagle’s Medium (Knockout DMEM) Thermo Fisher Scientific Cat# 10829018 Knockout Serum Replacement (KSR) Thermo Fisher Scientific Cat# 10828028 L-Glutamine Biological Industries Cat# 03-020-1A Nonessential amino acids Biological Industries Cat# 01-340-1B Penicillin and streptomycin Biological Industries Cat# 03-031-1B b-mercaptoethanol Sigma Cat# M7522 Human basic fibroblast growth factor (bFGF) PeproTech Cat# AF-100-18B Gelatin MP Biomedicals Cat# 960317 Fetal bovine serum (FBS) Thermo Fisher Scientific Cat# 10270106 polyethylenimine ‘‘Max’’ (PEI-Max) Polysciences Cat#24765 Trypsin–EDTA Biological Industries Cat# 03-052-1A Polybrene Sigma Cat# TR-1003 Puromycin dihydrochloride Sigma Cat# P8833 Blasticidin S hydrochloride Santa Cruz Biotechnology Cat# 3513-03-9 Matrigel Growth Factor Reduced (GFR) Corning Cat# 356231 Matrigel hESC-Qualified Matrix Corning Cat# 354277 SB431542 Biogems Cat# 3014193 LDN193189 Biogems Cat# 1062443 Dulbecco’s Modified Eagle’s Medium/ Nutrient Mixture F-12 Ham (DMEM/F12) Sigma Cat# D6421 N-2 supplement Thermo Fisher Cat# 17502001 Ascorbic acid Sigma Cat# A4544 D-Glucose BDH Cat# 10117 16% PFA Electron Microscopy Sciences Cat# 15710 DPBS Sigma-Aldrich Cat# D8662 Bovine serum albumin (BSA) WVR Cat# 0332 Donkey serum Sigma Cat# D9663 Triton X-100 Sigma-Aldrich Cat# T8787 Critical Commercial Assays STEMdiffTM Definitive Endoderm Kit STEMCELL Technologies Cat# 05110 STEMdiffTM Mesoderm Induction Kit STEMCELL Technologies Cat# 05220 Blood & Cell Culture DNA Midi Kit QIAGEN Cat# 13343 Nextera XT Index Kit, 96 indices, 384 samples Illumina Cat# FC-131-1002 (Continued on next page) e1 Cell Stem Cell 27, 1–13.e1–e6, October 1, 2020
Knock-Out:Article Title: Mapping Gene Circuits Essential for Germ Layer Differentiation via Loss-of-Function Screens in Haploid Human Embryonic Stem Cells.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies PerCP-Cy5.5-conjugated mouse anti-human SOX1 BD Biosciences Cat# 561549; RRID:AB_10694879 Alexa Flour 488-conjugated SOX17 BD Biosciences Cat# 562205; RRID:AB_10893402 goat anti-human/mouse NCAM1 R&D Systems Cat# AF2408; RRID: AB_442152 donkey anti-goat Cy5-conjugated secondary antibody Jackson ImmunoResearch Laboratories Cat# 705-175-147; RRID: AB_2340415 Chemicals, Peptides, and Recombinant Proteins mTeSR1 medium STEMCELL Technologies Cat# 05850 Y-27632-dihydrochloride Gentaur Cat# A3008 TrypLE Select Thermo Fisher Scientific Cat# 12563029 Knockout Dulbecco’s Modified Eagle’s Medium (Knockout DMEM) Thermo Fisher Scientific Cat# 10829018 Knockout Serum Replacement (KSR) Thermo Fisher Scientific Cat# 10828028 L-Glutamine Biological Industries Cat# 03-020-1A Nonessential amino acids Biological Industries Cat# 01-340-1B Penicillin and streptomycin Biological Industries Cat# 03-031-1B b-mercaptoethanol Sigma Cat# M7522 Human basic fibroblast growth factor (bFGF) PeproTech Cat# AF-100-18B Gelatin MP Biomedicals Cat# 960317 Fetal bovine serum (FBS) Thermo Fisher Scientific Cat# 10270106 polyethylenimine ‘‘Max’’ (PEI-Max) Polysciences Cat#24765 Trypsin–EDTA Biological Industries Cat# 03-052-1A Polybrene Sigma Cat# TR-1003 Puromycin dihydrochloride Sigma Cat# P8833 Blasticidin S hydrochloride Santa Cruz Biotechnology Cat# 3513-03-9 Matrigel Growth Factor Reduced (GFR) Corning Cat# 356231 Matrigel hESC-Qualified Matrix Corning Cat# 354277 SB431542 Biogems Cat# 3014193 LDN193189 Biogems Cat# 1062443 Dulbecco’s Modified Eagle’s Medium/ Nutrient Mixture F-12 Ham (DMEM/F12) Sigma Cat# D6421 N-2 supplement Thermo Fisher Cat# 17502001 Ascorbic acid Sigma Cat# A4544 D-Glucose BDH Cat# 10117 16% PFA Electron Microscopy Sciences Cat# 15710 DPBS Sigma-Aldrich Cat# D8662 Bovine serum albumin (BSA) WVR Cat# 0332 Donkey serum Sigma Cat# D9663 Triton X-100 Sigma-Aldrich Cat# T8787 Critical Commercial Assays STEMdiffTM Definitive Endoderm Kit STEMCELL Technologies Cat# 05110 STEMdiffTM Mesoderm Induction Kit STEMCELL Technologies Cat# 05220 Blood & Cell Culture DNA Midi Kit QIAGEN Cat# 13343 Nextera XT Index Kit, 96 indices, 384 samples Illumina Cat# FC-131-1002 (Continued on next page) e1 Cell Stem Cell 27, 1–13.e1–e6, October 1, 2020
Modification:Article Title: Mapping Gene Circuits Essential for Germ Layer Differentiation via Loss-of-Function Screens in Haploid Human Embryonic Stem Cells.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies PerCP-Cy5.5-conjugated mouse anti-human SOX1 BD Biosciences Cat# 561549; RRID:AB_10694879 Alexa Flour 488-conjugated SOX17 BD Biosciences Cat# 562205; RRID:AB_10893402 goat anti-human/mouse NCAM1 R&D Systems Cat# AF2408; RRID: AB_442152 donkey anti-goat Cy5-conjugated secondary antibody Jackson ImmunoResearch Laboratories Cat# 705-175-147; RRID: AB_2340415 Chemicals, Peptides, and Recombinant Proteins mTeSR1 medium STEMCELL Technologies Cat# 05850 Y-27632-dihydrochloride Gentaur Cat# A3008 TrypLE Select Thermo Fisher Scientific Cat# 12563029 Knockout Dulbecco’s Modified Eagle’s Medium (Knockout DMEM) Thermo Fisher Scientific Cat# 10829018 Knockout Serum Replacement (KSR) Thermo Fisher Scientific Cat# 10828028 L-Glutamine Biological Industries Cat# 03-020-1A Nonessential amino acids Biological Industries Cat# 01-340-1B Penicillin and streptomycin Biological Industries Cat# 03-031-1B b-mercaptoethanol Sigma Cat# M7522 Human basic fibroblast growth factor (bFGF) PeproTech Cat# AF-100-18B Gelatin MP Biomedicals Cat# 960317 Fetal bovine serum (FBS) Thermo Fisher Scientific Cat# 10270106 polyethylenimine ‘‘Max’’ (PEI-Max) Polysciences Cat#24765 Trypsin–EDTA Biological Industries Cat# 03-052-1A Polybrene Sigma Cat# TR-1003 Puromycin dihydrochloride Sigma Cat# P8833 Blasticidin S hydrochloride Santa Cruz Biotechnology Cat# 3513-03-9 Matrigel Growth Factor Reduced (GFR) Corning Cat# 356231 Matrigel hESC-Qualified Matrix Corning Cat# 354277 SB431542 Biogems Cat# 3014193 LDN193189 Biogems Cat# 1062443 Dulbecco’s Modified Eagle’s Medium/ Nutrient Mixture F-12 Ham (DMEM/F12) Sigma Cat# D6421 N-2 supplement Thermo Fisher Cat# 17502001 Ascorbic acid Sigma Cat# A4544 D-Glucose BDH Cat# 10117 16% PFA Electron Microscopy Sciences Cat# 15710 DPBS Sigma-Aldrich Cat# D8662 Bovine serum albumin (BSA) WVR Cat# 0332 Donkey serum Sigma Cat# D9663 Triton X-100 Sigma-Aldrich Cat# T8787 Critical Commercial Assays STEMdiffTM Definitive Endoderm Kit STEMCELL Technologies Cat# 05110 STEMdiffTM Mesoderm Induction Kit STEMCELL Technologies Cat# 05220 Blood & Cell Culture DNA Midi Kit QIAGEN Cat# 13343 Nextera XT Index Kit, 96 indices, 384 samples Illumina Cat# FC-131-1002 (Continued on next page) e1 Cell Stem Cell 27, 1–13.e1–e6, October 1, 2020
Electron Microscopy:Article Title: Mapping Gene Circuits Essential for Germ Layer Differentiation via Loss-of-Function Screens in Haploid Human Embryonic Stem Cells.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies PerCP-Cy5.5-conjugated mouse anti-human SOX1 BD Biosciences Cat# 561549; RRID:AB_10694879 Alexa Flour 488-conjugated SOX17 BD Biosciences Cat# 562205; RRID:AB_10893402 goat anti-human/mouse NCAM1 R&D Systems Cat# AF2408; RRID: AB_442152 donkey anti-goat Cy5-conjugated secondary antibody Jackson ImmunoResearch Laboratories Cat# 705-175-147; RRID: AB_2340415 Chemicals, Peptides, and Recombinant Proteins mTeSR1 medium STEMCELL Technologies Cat# 05850 Y-27632-dihydrochloride Gentaur Cat# A3008 TrypLE Select Thermo Fisher Scientific Cat# 12563029 Knockout Dulbecco’s Modified Eagle’s Medium (Knockout DMEM) Thermo Fisher Scientific Cat# 10829018 Knockout Serum Replacement (KSR) Thermo Fisher Scientific Cat# 10828028 L-Glutamine Biological Industries Cat# 03-020-1A Nonessential amino acids Biological Industries Cat# 01-340-1B Penicillin and streptomycin Biological Industries Cat# 03-031-1B b-mercaptoethanol Sigma Cat# M7522 Human basic fibroblast growth factor (bFGF) PeproTech Cat# AF-100-18B Gelatin MP Biomedicals Cat# 960317 Fetal bovine serum (FBS) Thermo Fisher Scientific Cat# 10270106 polyethylenimine ‘‘Max’’ (PEI-Max) Polysciences Cat#24765 Trypsin–EDTA Biological Industries Cat# 03-052-1A Polybrene Sigma Cat# TR-1003 Puromycin dihydrochloride Sigma Cat# P8833 Blasticidin S hydrochloride Santa Cruz Biotechnology Cat# 3513-03-9 Matrigel Growth Factor Reduced (GFR) Corning Cat# 356231 Matrigel hESC-Qualified Matrix Corning Cat# 354277 SB431542 Biogems Cat# 3014193 LDN193189 Biogems Cat# 1062443 Dulbecco’s Modified Eagle’s Medium/ Nutrient Mixture F-12 Ham (DMEM/F12) Sigma Cat# D6421 N-2 supplement Thermo Fisher Cat# 17502001 Ascorbic acid Sigma Cat# A4544 D-Glucose BDH Cat# 10117 16% PFA Electron Microscopy Sciences Cat# 15710 DPBS Sigma-Aldrich Cat# D8662 Bovine serum albumin (BSA) WVR Cat# 0332 Donkey serum Sigma Cat# D9663 Triton X-100 Sigma-Aldrich Cat# T8787 Critical Commercial Assays STEMdiffTM Definitive Endoderm Kit STEMCELL Technologies Cat# 05110 STEMdiffTM Mesoderm Induction Kit STEMCELL Technologies Cat# 05220 Blood & Cell Culture DNA Midi Kit QIAGEN Cat# 13343 Nextera XT Index Kit, 96 indices, 384 samples Illumina Cat# FC-131-1002 (Continued on next page) e1 Cell Stem Cell 27, 1–13.e1–e6, October 1, 2020
Cell Culture:Article Title: Mapping Gene Circuits Essential for Germ Layer Differentiation via Loss-of-Function Screens in Haploid Human Embryonic Stem Cells.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies PerCP-Cy5.5-conjugated mouse anti-human SOX1 BD Biosciences Cat# 561549; RRID:AB_10694879 Alexa Flour 488-conjugated SOX17 BD Biosciences Cat# 562205; RRID:AB_10893402 goat anti-human/mouse NCAM1 R&D Systems Cat# AF2408; RRID: AB_442152 donkey anti-goat Cy5-conjugated secondary antibody Jackson ImmunoResearch Laboratories Cat# 705-175-147; RRID: AB_2340415 Chemicals, Peptides, and Recombinant Proteins mTeSR1 medium STEMCELL Technologies Cat# 05850 Y-27632-dihydrochloride Gentaur Cat# A3008 TrypLE Select Thermo Fisher Scientific Cat# 12563029 Knockout Dulbecco’s Modified Eagle’s Medium (Knockout DMEM) Thermo Fisher Scientific Cat# 10829018 Knockout Serum Replacement (KSR) Thermo Fisher Scientific Cat# 10828028 L-Glutamine Biological Industries Cat# 03-020-1A Nonessential amino acids Biological Industries Cat# 01-340-1B Penicillin and streptomycin Biological Industries Cat# 03-031-1B b-mercaptoethanol Sigma Cat# M7522 Human basic fibroblast growth factor (bFGF) PeproTech Cat# AF-100-18B Gelatin MP Biomedicals Cat# 960317 Fetal bovine serum (FBS) Thermo Fisher Scientific Cat# 10270106 polyethylenimine ‘‘Max’’ (PEI-Max) Polysciences Cat#24765 Trypsin–EDTA Biological Industries Cat# 03-052-1A Polybrene Sigma Cat# TR-1003 Puromycin dihydrochloride Sigma Cat# P8833 Blasticidin S hydrochloride Santa Cruz Biotechnology Cat# 3513-03-9 Matrigel Growth Factor Reduced (GFR) Corning Cat# 356231 Matrigel hESC-Qualified Matrix Corning Cat# 354277 SB431542 Biogems Cat# 3014193 LDN193189 Biogems Cat# 1062443 Dulbecco’s Modified Eagle’s Medium/ Nutrient Mixture F-12 Ham (DMEM/F12) Sigma Cat# D6421 N-2 supplement Thermo Fisher Cat# 17502001 Ascorbic acid Sigma Cat# A4544 D-Glucose BDH Cat# 10117 16% PFA Electron Microscopy Sciences Cat# 15710 DPBS Sigma-Aldrich Cat# D8662 Bovine serum albumin (BSA) WVR Cat# 0332 Donkey serum Sigma Cat# D9663 Triton X-100 Sigma-Aldrich Cat# T8787 Critical Commercial Assays STEMdiffTM Definitive Endoderm Kit STEMCELL Technologies Cat# 05110 STEMdiffTM Mesoderm Induction Kit STEMCELL Technologies Cat# 05220 Blood & Cell Culture DNA Midi Kit QIAGEN Cat# 13343 Nextera XT Index Kit, 96 indices, 384 samples Illumina Cat# FC-131-1002 (Continued on next page) e1 Cell Stem Cell 27, 1–13.e1–e6, October 1, 2020
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