ProductHighlights:
TheIlluminaBio-Rad®SureCell™WholeTranscriptomeAnalysis(WTA)3’Kitprovidesavalidatedprocessforsingle-cellRNAsequencing.
- Sensitiveandunbiasedcharacterizationoftranscriptionalsignatures–HighlysensitiveandreproducIBLegenedetectioninvariedindividualcells
- Comprehensivesingle-cellRNAsequencingworkflow–Fullysupportedworkflowdevelopedincollaborationbytechnologyinnovators
- Powerfulnext-generationsequencingintegratedwithsimpledataanalysis–ProvenIlluminasequencingcombinedwithstreamlined,user-friendlyanalysissoftware
Singlecellsareindividuallypartitionedintosubnanoliterdropletsonadisposablecartridgeontheone-touchddSEQ™Single-CellIsolator.Thecartridgecanaccommodatemultiplesamples,andmultiplecartridgescanbeprocessedinparalleltoisolatethousandsofcellsinamatterofminutes.
Celllysisandcellbarcodingoccurinsideindividualdroplets,andsingle-cell-barcodedRNA-SeqlibrariesaresubsequentlypreparedusingNexteratechnology.
DataAnalysis
ConductdataanalysisviaBaseSpaceSequenceHub,theIlluminacloud-basedgenomicscomputingenvironment.
SpeciesDetails
Thiskithasbeenvalidatedforhumanandmousesamples.TheBaseSpaceSingle-CellRNA-SeqAppsupportsanalysisofhuman,mouse,rat,zebrafish,C.elegans,andDrosophilasamples.
Specifications:
| SystemCompatibility | NextSeq550,HiSeq2000,HiSeq3000,HiSeq1000,HiSeq1500,NextSeq500,HiSeq2500,NovaSeq6000,HiSeq4000 |
| SpeciesCategory | Human,Mouse |
| ContentSpecifications | Capturesthecodingtranscriptome(3u0027enriched),withstrand-specificinformation |
| Method | Whole-TranscriptomeSequencing,mRNASequencing |
| Technology | Sequencing |
| SpecializedSampleTypes | SingleCells |
Unbiased,Cell-SizeAgnosticRNA-Seq:
Mousecelllines(A20,NIH3T3)andhumancelllines(HEK,BJ)wereprocessedusingtheSureCellWTA3′LibraryPrepKit.Consistentlyhighnumbersofdetectedgenesdemonstratesthatrecoveryoftranscriptsisnotlimitedbycellsize.ConfidentSingle-CellTranscriptIdentification:
A.BaseSpace-generatedplotsofthenumberofuniquemolecularidentifiers(UMI),i.e.transcripts,assignedtothemouse/mm10(red)andhuman/hg19(blue)genomesforeachcellbarcode.Uniquetranscriptsmappingtobothmouseandhuman(purple)representcelldoublets.B.Cumulatedfractionofuniquetranscriptsassignedtocellbarcodes(linearscale).Theinflectionpoint(redline)determinesthenumberofbarcodedcellsdetectedinthesequencingrun.ConfidentCellPopulationIdentification:
A.PCAclusteringof1384singlecellssortedfroma1:1ratiomixtureofHEK293andNIH3T3cells,sequencedandanalyzedwiththeSingle-CellRNA-SeqBaseSpaceApp.B.Cellpopulationsclusterbasedonexpressionofthehuman(hg19)RPL13gene.ConfidentCellSubpopulationIdentification:
A.AnalysisintheSingle-CellRNA-SeqBaseSpaceAppusingthet-SNEalgorithmofamixtureofNIH3T3andHEK293cellsidentifiesadistinctsubpopulationofcells.B.Cellscolorcodedbygeneexpressionofhg19RPL13confirmstheidentityofthesubpopulationashuman.CellCycleAnalysisofSingleCellsbyRNA-Seq:
AnalysisofcellcyclestateusingtheSingle-CellRNA-SeqBaseSpaceAppisbasedonuniquetranscriptcountsofgenesassociatedwitheachphaseofthecellcycle,normalizedbythetotalcountforeachcell.Expressioniscenteredbythemedianandscaledbythemedianabsolutedeviationforeachcellcycle.
