{"id":"https://openalex.org/W3082607384","doi":"https://doi.org/10.17504/protocols.io.sh4eb8w","title":"G418 Kill curve protocol v1","display_name":"G418 Kill curve protocol v1","publication_year":2018,"publication_date":"2018-08-09","ids":{"openalex":"https://openalex.org/W3082607384","doi":"https://doi.org/10.17504/protocols.io.sh4eb8w","mag":"3082607384"},"language":"en","primary_location":{"id":"doi:10.17504/protocols.io.sh4eb8w","doi":"https://doi.org/10.17504/protocols.io.sh4eb8w","is_oa":true,"landing_page_url":"https://doi.org/10.17504/protocols.io.sh4eb8w","pdf_url":"https://www.protocols.io/view/g418-kill-curve-protocol-sh4eb8w.pdf","source":null,"license":"cc-by","license_id":"https://openalex.org/licenses/cc-by","version":"acceptedVersion","is_accepted":true,"is_published":false,"raw_source_name":null,"raw_type":"posted-content"},"volume":null,"type":"preprint","indexed_in":["crossref"],"open_access":{"is_oa":true,"oa_status":"green","oa_url":"https://www.protocols.io/view/g418-kill-curve-protocol-sh4eb8w.pdf","any_repository_has_fulltext":null},"authorships":[{"author_position":"first","author":{"id":"https://openalex.org/A5056393791","display_name":"Fatma Gomaa","orcid":"https://orcid.org/0000-0001-5697-7118","observed_orcids":["https://orcid.org/0000-0001-5697-7118"]},"institutions":[{"id":"https://openalex.org/I136199984","display_name":"Harvard University","ror":"https://ror.org/03vek6s52","country_code":"US","type":"education","lineage":["https://openalex.org/I136199984"]},{"id":"https://openalex.org/I165733156","display_name":"University of Georgia","ror":"https://ror.org/00te3t702","country_code":"US","type":"education","lineage":["https://openalex.org/I165733156"]},{"id":"https://openalex.org/I66958751","display_name":"Woods Hole Oceanographic Institution","ror":"https://ror.org/03zbnzt98","country_code":"US","type":"funder","lineage":["https://openalex.org/I66958751"]}],"countries":["US"],"is_corresponding":false,"raw_author_name":"Fatma Gomaa","raw_affiliation_strings":["Harvard University","Harvard University ; University of Georgia ; Woods Hole Oceanographic Institution Protist Research to Optimize Tools in Genetics (PROT-G)","Harvard University; University of Georgia; Woods Hole Oceanographic Institution Protist Research"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Harvard University","institution_ids":["https://openalex.org/I136199984"]},{"raw_affiliation_string":"Harvard University ; University of Georgia ; Woods Hole Oceanographic Institution Protist Research to Optimize Tools in Genetics (PROT-G)","institution_ids":["https://openalex.org/I136199984","https://openalex.org/I165733156","https://openalex.org/I66958751"]},{"raw_affiliation_string":"Harvard University; University of Georgia; Woods Hole Oceanographic Institution Protist Research","institution_ids":["https://openalex.org/I136199984","https://openalex.org/I165733156","https://openalex.org/I66958751"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5033840058","display_name":"Zhu\u2010Hong Li","orcid":"https://orcid.org/0000-0003-4940-0729","observed_orcids":["https://orcid.org/0000-0003-4940-0729"]},"institutions":[{"id":"https://openalex.org/I136199984","display_name":"Harvard University","ror":"https://ror.org/03vek6s52","country_code":"US","type":"education","lineage":["https://openalex.org/I136199984"]},{"id":"https://openalex.org/I165733156","display_name":"University of Georgia","ror":"https://ror.org/00te3t702","country_code":"US","type":"education","lineage":["https://openalex.org/I165733156"]},{"id":"https://openalex.org/I66958751","display_name":"Woods Hole Oceanographic Institution","ror":"https://ror.org/03zbnzt98","country_code":"US","type":"funder","lineage":["https://openalex.org/I66958751"]}],"countries":["US"],"is_corresponding":false,"raw_author_name":"Zhuhong Li","raw_affiliation_strings":["Harvard University ; University of Georgia ; Woods Hole Oceanographic Institution Protist Research to Optimize Tools in Genetics (PROT-G)","Harvard University; University of Georgia; Woods Hole Oceanographic Institution Protist Research"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Harvard University ; University of Georgia ; Woods Hole Oceanographic Institution Protist Research to Optimize Tools in Genetics (PROT-G)","institution_ids":["https://openalex.org/I136199984","https://openalex.org/I165733156","https://openalex.org/I66958751"]},{"raw_affiliation_string":"Harvard University; University of Georgia; Woods Hole Oceanographic Institution Protist Research","institution_ids":["https://openalex.org/I136199984","https://openalex.org/I165733156","https://openalex.org/I66958751"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5000842323","display_name":"Roberto Docampo","orcid":"https://orcid.org/0000-0003-4229-8784","observed_orcids":["https://orcid.org/0000-0003-4229-8784"]},"institutions":[{"id":"https://openalex.org/I136199984","display_name":"Harvard University","ror":"https://ror.org/03vek6s52","country_code":"US","type":"education","lineage":["https://openalex.org/I136199984"]},{"id":"https://openalex.org/I165733156","display_name":"University of Georgia","ror":"https://ror.org/00te3t702","country_code":"US","type":"education","lineage":["https://openalex.org/I165733156"]},{"id":"https://openalex.org/I66958751","display_name":"Woods Hole Oceanographic Institution","ror":"https://ror.org/03zbnzt98","country_code":"US","type":"funder","lineage":["https://openalex.org/I66958751"]}],"countries":["US"],"is_corresponding":false,"raw_author_name":"Roberto Docampo","raw_affiliation_strings":["Harvard University ; University of Georgia ; Woods Hole Oceanographic Institution Protist Research to Optimize Tools in Genetics (PROT-G)","Harvard University; University of Georgia; Woods Hole Oceanographic Institution Protist Research"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Harvard University ; University of Georgia ; Woods Hole Oceanographic Institution Protist Research to Optimize Tools in Genetics (PROT-G)","institution_ids":["https://openalex.org/I136199984","https://openalex.org/I165733156","https://openalex.org/I66958751"]},{"raw_affiliation_string":"Harvard University; University of Georgia; Woods Hole Oceanographic Institution Protist Research","institution_ids":["https://openalex.org/I136199984","https://openalex.org/I165733156","https://openalex.org/I66958751"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5048349346","display_name":"Peter R. Girguis","orcid":"https://orcid.org/0000-0002-3599-8160","observed_orcids":["https://orcid.org/0000-0002-3599-8160","https://orcid.org/0000-0001-9006-5411"]},"institutions":[{"id":"https://openalex.org/I136199984","display_name":"Harvard University","ror":"https://ror.org/03vek6s52","country_code":"US","type":"education","lineage":["https://openalex.org/I136199984"]},{"id":"https://openalex.org/I165733156","display_name":"University of Georgia","ror":"https://ror.org/00te3t702","country_code":"US","type":"education","lineage":["https://openalex.org/I165733156"]},{"id":"https://openalex.org/I66958751","display_name":"Woods Hole Oceanographic Institution","ror":"https://ror.org/03zbnzt98","country_code":"US","type":"funder","lineage":["https://openalex.org/I66958751"]}],"countries":["US"],"is_corresponding":false,"raw_author_name":"Peter Girguis","raw_affiliation_strings":["Harvard University ; University of Georgia ; Woods Hole Oceanographic Institution Protist Research to Optimize Tools in Genetics (PROT-G)","Harvard University; University of Georgia; Woods Hole Oceanographic Institution Protist Research"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Harvard University ; University of Georgia ; Woods Hole Oceanographic Institution Protist Research to Optimize Tools in Genetics (PROT-G)","institution_ids":["https://openalex.org/I136199984","https://openalex.org/I165733156","https://openalex.org/I66958751"]},{"raw_affiliation_string":"Harvard University; University of Georgia; Woods Hole Oceanographic Institution Protist Research","institution_ids":["https://openalex.org/I136199984","https://openalex.org/I165733156","https://openalex.org/I66958751"]}]},{"author_position":"last","author":{"id":"https://openalex.org/A5020626624","display_name":"Virginia P. Edgcomb","orcid":"https://orcid.org/0000-0001-6805-381X","observed_orcids":["https://orcid.org/0000-0001-6805-381X"]},"institutions":[{"id":"https://openalex.org/I136199984","display_name":"Harvard University","ror":"https://ror.org/03vek6s52","country_code":"US","type":"education","lineage":["https://openalex.org/I136199984"]},{"id":"https://openalex.org/I165733156","display_name":"University of Georgia","ror":"https://ror.org/00te3t702","country_code":"US","type":"education","lineage":["https://openalex.org/I165733156"]},{"id":"https://openalex.org/I66958751","display_name":"Woods Hole Oceanographic Institution","ror":"https://ror.org/03zbnzt98","country_code":"US","type":"funder","lineage":["https://openalex.org/I66958751"]}],"countries":["US"],"is_corresponding":false,"raw_author_name":"Virginia Edgcomb","raw_affiliation_strings":["Harvard University ; University of Georgia ; Woods Hole Oceanographic Institution Protist Research to Optimize Tools in Genetics (PROT-G)","Harvard University; University of Georgia; Woods Hole Oceanographic Institution Protist Research"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Harvard University ; University of Georgia ; Woods Hole Oceanographic Institution Protist Research to Optimize Tools in Genetics (PROT-G)","institution_ids":["https://openalex.org/I136199984","https://openalex.org/I165733156","https://openalex.org/I66958751"]},{"raw_affiliation_string":"Harvard University; University of Georgia; Woods Hole Oceanographic Institution Protist Research","institution_ids":["https://openalex.org/I136199984","https://openalex.org/I165733156","https://openalex.org/I66958751"]}]}],"institutions":[],"countries_distinct_count":1,"institutions_distinct_count":3,"corresponding_author_ids":[],"corresponding_institution_ids":[],"apc_list":null,"apc_paid":null,"fwci":null,"has_fulltext":true,"cited_by_count":0,"citation_normalized_percentile":null,"cited_by_percentile_year":null,"biblio":{"volume":null,"issue":null,"first_page":null,"last_page":null},"is_retracted":false,"is_paratext":false,"is_xpac":false,"primary_topic":{"id":"https://openalex.org/T10120","display_name":"Bacterial Genetics and Biotechnology","score":0.510699987411499,"subfield":{"id":"https://openalex.org/subfields/1311","display_name":"Genetics"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},"topics":[{"id":"https://openalex.org/T10120","display_name":"Bacterial Genetics and Biotechnology","score":0.510699987411499,"subfield":{"id":"https://openalex.org/subfields/1311","display_name":"Genetics"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},{"id":"https://openalex.org/T10497","display_name":"Fungal and yeast genetics research","score":0.04610000178217888,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},{"id":"https://openalex.org/T11663","display_name":"Viral Infectious Diseases and Gene Expression in Insects","score":0.033399999141693115,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}}],"keywords":[{"id":"https://openalex.org/keywords/bodo-saltans","display_name":"Bodo saltans","score":0.8920000195503235},{"id":"https://openalex.org/keywords/antibiotic-selection","display_name":"antibiotic selection","score":0.5669999718666077},{"id":"https://openalex.org/keywords/neomycin-resistance-gene","display_name":"neomycin resistance gene","score":0.9240000247955322},{"id":"https://openalex.org/keywords/transfection","display_name":"transfection","score":0.5690000057220459},{"id":"https://openalex.org/keywords/electroporation","display_name":"electroporation","score":0.824999988079071},{"id":"https://openalex.org/keywords/cell-viability","display_name":"cell viability","score":0.35600000619888306}],"concepts":[{"id":"https://openalex.org/C19374876","wikidata":"https://www.wikidata.org/wiki/Q247170","display_name":"Hemocytometer","level":2,"score":0.6842246651649475},{"id":"https://openalex.org/C54009773","wikidata":"https://www.wikidata.org/wiki/Q1429031","display_name":"Transfection","level":3,"score":0.5323373079299927},{"id":"https://openalex.org/C81885089","wikidata":"https://www.wikidata.org/wiki/Q189082","display_name":"Cell culture","level":2,"score":0.4935716390609741},{"id":"https://openalex.org/C86803240","wikidata":"https://www.wikidata.org/wiki/Q420","display_name":"Biology","level":0,"score":0.4866279065608978},{"id":"https://openalex.org/C29730261","wikidata":"https://www.wikidata.org/wiki/Q274160","display_name":"Toxicity","level":2,"score":0.48180001974105835},{"id":"https://openalex.org/C7623868","wikidata":"https://www.wikidata.org/wiki/Q1967135","display_name":"Centrifugation","level":2,"score":0.44885537028312683},{"id":"https://openalex.org/C153911025","wikidata":"https://www.wikidata.org/wiki/Q7202","display_name":"Molecular biology","level":1,"score":0.44781920313835144},{"id":"https://openalex.org/C2779841869","wikidata":"https://www.wikidata.org/wiki/Q423098","display_name":"Neomycin","level":3,"score":0.42304527759552},{"id":"https://openalex.org/C185592680","wikidata":"https://www.wikidata.org/wiki/Q2329","display_name":"Chemistry","level":0,"score":0.31528183817863464},{"id":"https://openalex.org/C89423630","wikidata":"https://www.wikidata.org/wiki/Q7193","display_name":"Microbiology","level":1,"score":0.2343761920928955},{"id":"https://openalex.org/C501593827","wikidata":"https://www.wikidata.org/wiki/Q12187","display_name":"Antibiotics","level":2,"score":0.2242279052734375},{"id":"https://openalex.org/C55493867","wikidata":"https://www.wikidata.org/wiki/Q7094","display_name":"Biochemistry","level":1,"score":0.20058917999267578},{"id":"https://openalex.org/C54355233","wikidata":"https://www.wikidata.org/wiki/Q7162","display_name":"Genetics","level":1,"score":0.10892641544342041},{"id":"https://openalex.org/C178790620","wikidata":"https://www.wikidata.org/wiki/Q11351","display_name":"Organic chemistry","level":1,"score":0.0}],"mesh":[],"locations_count":1,"locations":[{"id":"doi:10.17504/protocols.io.sh4eb8w","doi":"https://doi.org/10.17504/protocols.io.sh4eb8w","is_oa":true,"landing_page_url":"https://doi.org/10.17504/protocols.io.sh4eb8w","pdf_url":"https://www.protocols.io/view/g418-kill-curve-protocol-sh4eb8w.pdf","source":null,"license":"cc-by","license_id":"https://openalex.org/licenses/cc-by","version":"acceptedVersion","is_accepted":true,"is_published":false,"raw_source_name":null,"raw_type":"posted-content"}],"best_oa_location":{"id":"doi:10.17504/protocols.io.sh4eb8w","doi":"https://doi.org/10.17504/protocols.io.sh4eb8w","is_oa":true,"landing_page_url":"https://doi.org/10.17504/protocols.io.sh4eb8w","pdf_url":"https://www.protocols.io/view/g418-kill-curve-protocol-sh4eb8w.pdf","source":null,"license":"cc-by","license_id":"https://openalex.org/licenses/cc-by","version":"acceptedVersion","is_accepted":true,"is_published":false,"raw_source_name":null,"raw_type":"posted-content"},"sustainable_development_goals":[],"study_designs":[],"awards":[],"funders":[],"has_content":{"grobid_xml":true,"pdf":true},"content_urls":{"pdf":"https://content.openalex.org/works/W3082607384.pdf","grobid_xml":"https://content.openalex.org/works/W3082607384.grobid-xml"},"referenced_works_count":0,"referenced_works":[],"related_works":["https://openalex.org/W3012432682","https://openalex.org/W2789960100","https://openalex.org/W2408952450","https://openalex.org/W2006845914","https://openalex.org/W2521265097","https://openalex.org/W1994898104","https://openalex.org/W125860204","https://openalex.org/W2364350236","https://openalex.org/W2093666165","https://openalex.org/W2406196067"],"abstract_inverted_index":{"Bodo":[0],"saltans":[1,100,137,285,306],"kill":[2,96,281,349],"curve":[3,97,350],"protocol":[4],"using":[5,188,208],"G418":[6,29,86,104,194,216,276,302,341],"(Gentamicin)":[7],"Gentamycin":[8],"exhibits":[9],"toxicity":[10],"toward":[11],"both":[12],"eukaryotic":[13,39],"and":[14,68,119,363],"prokaryotic":[15],"cells":[16,40,62,82,226,243,262,299,337,365,372,382,386],"by":[17,53,147,403],"disrupting":[18],"ribosome":[19],"function,":[20],"thereby":[21],"blocking":[22],"the":[23,80,152,156,178,181,192,212,220,236,282,297,317,332,348,355,380],"elongation":[24],"step":[25],"in":[26,64,171,180,287],"protein":[27],"synthesis.":[28],"is":[30,51,140,219,235,254,258,291],"most":[31],"commonly":[32],"used":[33,71,125],"as":[34,78,397],"a":[35,45,197,251,255,275,398],"selection":[36,116,218],"agent":[37],"for":[38,99,115,126,201,217,223,295,304,366,408],"genetically":[41],"engineered":[42],"to":[43,72,85,112,261,323,383,388,406],"express":[44],"neomycin":[46],"resistance":[47],"gene":[48,319],"(NeoR),":[49],"which":[50,224,238,257,290],"encoded":[52],"either":[54],"transposon":[55],"Tn601":[56],"(903)":[57],"or":[58,395,401],"Tn5.":[59],"The":[60,107,271],"resistant":[61,298,321],"grow":[63,387],"medium":[65,158,161,179,190,334,357],"containing":[66,162],"G418,":[67],"may":[69,392],"be":[70,124,393],"establish":[73,95],"stably":[74],"transfected":[75,129,326,336,381],"cell":[76,130,206],"lines":[77],"all":[79,225,310],"non-resistant":[81],"die":[83,374],"due":[84],"toxicity,":[87,204],"typically":[88],"within":[89,263,375],"6":[90,174],"\u2013":[91,143,164,185,360],"14":[92],"days.":[93],"To":[94],"assay":[98],"various":[101],"concentrations":[102,122],"of":[103,117,203,215,247,266,278,311,335],"were":[105,338],"tested.":[106],"concentration":[108,237,256,277],"range":[109],"from":[110],"1":[111],"50":[113,165],"\u03bcg/mL":[114,280],"eukaryotic,":[118],"then":[120],"higher":[121],"can":[123],"maintaining":[127],"stable":[128],"line.":[131],"Kill":[132],"Curve":[133],"Assay":[134],"Harvestthe":[135],"B.":[136,284,305],"cellsfromaculture":[138],"that":[139,211,274],"atpeakdensity":[141],"(1":[142],"3.0":[144],"x":[145],"105)":[146],"centrifugation":[148],"at":[149],"800xgfor5min.,":[150],"discard":[151],"supernatant":[153],"(medium).":[154],"Replace":[155,354],"growth":[157,333],"with":[159,191,340],"fresh":[160,189],"0":[163],"\u03bcg/mL.":[166],"For":[167],"each":[168],"concentration,":[169],"test":[170],"triplicate.":[172],"Using":[173],"wells":[175,182],"plates,":[176],"replace":[177],"every":[183,358],"3":[184,359],"4":[186],"days":[187,265,289,362],"appropriate":[193],"concentration.":[195],"Perform":[196],"daily":[198],"visual":[199,367],"inspection":[200],"evidence":[202],"also":[205],"count":[207],"hemocytometer.":[209],"Note":[210],"optimal":[213],"dose":[214,234,253],"lowest":[221],"one":[222,230],"have":[227],"died":[228],"after":[229,244,300,330],"week.":[231],"A":[232],"low":[233],"has":[239],"minimal":[240],"effects":[241],"on":[242,347],"2":[245,279],"weeks":[246],"antibiotic":[248,268,342],"selection.":[249,269],"Conversely,":[250],"high":[252,389],"highly":[259],"toxic":[260],"2-3":[264],"starting":[267],"5-":[270],"results":[272,351],"indicate":[273],"entire":[283],"population":[286],"12":[288,377],"considered":[292],"enough":[293],"time":[294],"selecting":[296],"transfection.":[301],"Sensitivity":[303],"Cell":[307],"Selection":[308],"In":[309],"our":[312,325],"plasmids":[313],"constructs":[314],"we":[315],"included":[316],"NeoR":[318],"(Neomycine":[320],"gene)":[322],"select":[324,407],"cells.":[327],"Twenty-four":[328],"hours":[329],"electroporation,":[331],"supplemented":[339],"Solution":[343],"(2":[344],"\u03bcg/mL)":[345],"based":[346],"(above).":[352],"3.":[353],"G418-containing":[356],"5":[361],"examine":[364],"toxicity.":[368],"Most":[369],"non-transfected":[370],"(non-resistant)":[371],"will":[373],"10-":[376],"days,":[378],"leaving":[379],"expand.":[384],"Once":[385],"confluence,":[390],"they":[391],"maintained,":[394],"frozen":[396],"polyclonal":[399],"line":[400],"plated":[402],"limited":[404],"dilution":[405],"single":[409],"clones.":[410]},"counts_by_year":[],"updated_date":"2026-10-28T10:33:24.094632","created_date":"2020-09-08T00:00:00"}
