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.ebiomall.com
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便宜点的就是国产的tiangen,效果会差一点点 如果是大样本的全血 用这个也凑合
根据模板的类型分为DNA类PCR试剂盒和RNA类PCR试剂盒
PCR mix厂家:普洛麦格公司,QIAGEN公司,罗氏公司,TAKARA公司.
成品PCR试剂盒厂家:深圳匹基,上海之江,中山达安,上海科华,上海复星.
2、电泳问题:如果你核酸定量浓度不低的话就考虑电泳问题,电泳的时候加上1kb的marker,如果marker跑不出来说明你凝胶有问题,一般只要marker跑出来了,DNA浓度又比较高,而没有条带这样的现象很少见,DNA提取比较简单而且相当稳定,本人当时做甲基化提取的DNA有一次拿出来电泳忘了放回冰箱了,结果大夏天的在外面放了一天,心怀忐忑的进行了一次电泳,结果条带依然给力,一点都没有降解.
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