欢迎来到天天文库
浏览记录
ID:40382040
大小:673.03 KB
页数:41页
时间:2019-08-01
《Human mtDNA Genome Guide (15037958)》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、HumanmtDNAGenomeFortheIlluminaSequencingPlatformIntroduction2Workflow3SampleSheet4FirstPCRAmplification6Quantify9TagmentAmplifiedDNA11Limited‐CyclePCRAmplification13Limited‐CyclePCRClean‐Up17Bead‐basedNormalization19PoolandLoadfromBead‐basedNormalization23Bioanalyzer‐basedNormalization25Poola
2、ndLoadfromBioanalyzer‐basedNormalization27Analysis29SupportingInformation36IMPORTANTThisdocumentprovidesinformationforanapplicationforIlluminatechnologythathasbeendemonstratedinternallyandmaybeofinteresttocustomers.ThisinformationisNOTICEprovidedas‐isandisnotanIlluminaproductandisnotaccompani
3、edbyanyrightsorwarranties.Customersusingoradaptingthisinformationshouldobtainanylicensesrequiredandmaterialsfromauthorizedvendors.Illuminaproductsmentionedhereinareforresearchuseonlyunlessmarkedotherwise.Whilecustomerfeedbackiswelcomed,thisapplicationisnotsupportedbyIlluminaTechnicalSupportan
4、dFieldApplicationScientists.Part#15037958Rev.BPage1IntroductionPage2IntroductionThisprotocolexplainshowtoprepare,sequence,andanalyzetheentirehumanmitochondrialDNA(mtDNA)genomefromclean,intactDNAsamples.Duringsamplepreparation,themtDNAgenomeisamplifiedintwoPCRstogeneratetwolongfragmentsspannin
5、gtheentirehumanmitochondrialgenome(16,569bp).Theampliconsarequantifiedandpooledbeforelibrarypreparation.SubsequentlibrarysequencingontheMiSeqisfollowedbydataanalysiswithMiSeqReporterandvariantcallingwiththemtDNAAnalysisTool.NOTETheprotocolsdescribedinthisguideassumethatyouhavereviewedtheSuppo
6、rtingInformation,onpage36andobtainedalloftherequisiteconsumablesandequipment.What’sNewThefollowingchangesweremadeinthisrevision:•AsafestoppingpointwasaddedattheendoftheLimited‐CyclePCRClean‐Upprocedure.•TheNexteraXTDNASamplePreparationKit(96samples)andNexteraXTDNASamplePreparationIndexKitswer
7、eaddedtotheconsumableslist.WorkflowPage3WorkflowThefollowingfigureillustratestheHumanmtDNAGenomeworkflow.Figure1HumanmtDNAGenomeWorkflowSampleSheetPage4SampleSheetAftercreatingyoursamplesheetusingtheIlluminaExperimentManager(IEM),openthesampl
此文档下载收益归作者所有