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1、ANALYSISQuantitativeassessmentofsingle-cellRNA-sequencingmethodsAngelaRWu1,NormaFNeff1,TomerKalisky1,8,PieroDalerba2–4,BarbaraTreutlein1,MichaelERothenberg5,FrancisMMburu1,6,GaryLMantalas1,SopheakSim3,MichaelFClarke2–4&StephenRQuake1,6,7Interestinsingle-cellwhole-transcriptomeanalysisi
2、sHowever,inordertoassessthevalidityofameasurement,itgrowingrapidly,especiallyforprofilingrareorheterogeneousisalsocriticaltoevaluateaccuracy,orhowclosethemeasure-populationsofcells.Wecomparedcommerciallyavailablementcomestothetruevalue.Accuracydependsonsystematicsingle-cellRNAamplificati
3、onmethodswithbothmicrolitererrorsderivingfromthedatacollectionmethod,anditisoftenandnanolitervolumes,usingsequencefrombulktotalRNAandestimatedbyusingdifferentmeasurementtechniquesonthesamemultiplexedquantitativePCRasbenchmarkstosystematicallysampletype.evaluatethesensitivityandaccuracy
4、ofvarioussingle-cellHerewereportquantitativeRNA-seqanalysisof102single-cellRNA-seqapproaches.Weshowthatsingle-cellRNA-seqcanhumantranscriptomes.Weassessedtheperformanceofcom-beusedtoperformaccuratequantitativetranscriptomemerciallyavailablesingle-cellRNAamplificationmethodsinmeasuremen
5、tinindividualcellswitharelativelysmallnumberbothmicroliterandnanolitervolumes,comparedeachmethodtoofsequencingreadsandthatsequencinglargenumbersofconventionalRNA-seqofthesamesampleusingbulktotalRNAsinglecellscanrecapitulatebulktranscriptomecomplexity.andevaluatedtheaccuracyofthemeasure
6、mentsbyindepend-entlyquantitatingexpressionof40genesinthesamecelltypebyHigh-throughputsequencingofwholetranscriptomes,orRNA-multiplexedquantitativePCR(qPCR)16,17.Ourresultsshowthatseq,hasbeenusedextensivelytoprofilegeneexpressionfromitispossibletousesingle-cellRNA-seqtoperformquantitat
7、iveNatureAmerica,Inc.Allrightsreserved.bulktissues.Thereisagrowingdemandformethodsthatallowtranscriptomemeasurementsofsinglecellsandthat,whensuch4whole-transcriptomeprofilingofsinglecells,drivenby(i)themeasurementsareperformedonlargenumbersofcells,onecanneedfordirectanalysisofrarecel
