TheIonTorrentNGSReverseTranscription(RT)KitisthefirstcDNAsynthesiskitdevelopedspecificallyfornext-generationsequencing(NGS)applications.ItcombinesthesuperiorperformanceofSuperScriptIVReverseTranscriptasewithanovelmastermixoptimizedforNGSlibrarypreparation.TheNGSRTKitiscompatiblewithIonAmpliSeqandIonAmpliSeqHDassaysandcanbeusedwithbothmanualandautomatedworkflows.

BenefitsoftheNGSRTKitinclude:
Optimizedforavarietyofsampletypes,includingFFPE,degradedsamples,andthosewithlowtemplateamounts
SuperiorNGSresultswithnovelRTformulationoptimizedforNGSlibrarypreparation
HelpsincreaseNGSlibraryyield,improvedetectionoflongamplicons,andrescueunder-performingamplicons
HelpsmaximizemappedreadsandimprovedetectionofRNAfusions
TrustedSuperScriptIVRTtechnologyofferssuperiorcDNAsynthesisperformancewitheventhemostchallengingRNAsamples
UniversalkitforwidevarietyofOncomineoncologyresearchassaysandIonAmpliSeqpanels

Simplereactionsetup
TheNGSRTKitcontainsatubeof10XSuperScriptIVenzymemixandatubeof5Xbuffer.Aone-step,25-minutereactionreversetranscribestotalRNAintocDNAandcanbeuseddirectlyforNGSlibrarypreparation,withoutanyadditionalquantification,purification,ordilutionsteps.Avarietyofsampletypescanbeused,includingRNAfromblood,bonemarrow,FFPEandpartiallydegradedsamples.ThisproductiscompatiblewithOncomineandIonAmpliSeqworkflows,aswellasIonAmpliSeqHDpanels.ConsultthecDNAsynthesissectionofeachassaymanualforspecificRTreactionvolumesandsetupinstructions.

Performanceandqualitytesting
TheNGSRTKitisassayedforendodeoxyribonuclease,exodeoxyribonuclease,andribonuclease,aswellasyieldandlengthofcDNAproduct.ItisQCtestedwithafullNGSworkflowtoevaluatelibraryyieldandensureperformancemeetskeysequencingmetrics.


TheAppliedBiosystemsTaqManRNA-to-CT1-StepKitdeliversconsistentRNAtargetquantificationforawidevarietyofassaysandisvalidatedwithAppliedBiosystemscomprehensivelibraryofTaqManGeneExpressionassays.Thekitisrecommendedforavarietyofapplicationsincludinggeneralgeneexpressionstudies,biomarkerdiscovery,andmicroarrayvalidation.

FeaturesoftheTaqManRNA-to-CT1-StepKitinclude:
•Accurateacrossawidedynamicrangefordependableperformance
•ValidatedwithTaqManGeneExpressionAssaysforeasyexperimentalsetup
•Consistentperformancewithawidevarietyoftargets,includingAT-rich,GC-richandlongsequences
•1-Stepformatmeansfewerpipettingstepsleadingtolesschancesforerrors

Efficiencyacrossabroadrangeofinputconcentrations
MaximumflexibilityinRNAtemplateinputquantityrequiresoptimalreal-timePCRefficiencyacrossabroadrangeoftemplatequantities.Figure1showstheexcellentamplificationefficiencyacrosssixordersofmagnitude.TheresultiscomparabletothatoftheTaqManRNA-to-CT2-StepKit,andisasignificantimprovementindynamicrangeandsensitivityovertheTaqManOne-StepRT-PCRMasterMixReagents.

Highsensitivityatlowtargetconcentration
ThesensitivityofTaqManRNA-to-CT1-StepKitwasvalidatedusingasyntheticRNAtemplateforwhichcopynumberispreciselyknown.Significantsamplingerroroccurswhenmeasuringlowquantitiesoftarget,sostatisticalanalysisisrequiredforproperevaluationusingmultiplereplicates.Figure2showstheexpectedquantityoftargetandcorrespondingmeanCTvalues.StatisticalanalysisindicateshighconfidenceofsamplequantificationbasedonaT-test(TableI),consistentwithasfewas10copiesoftarget*.

*Resultsaredependentonavarietyoffactors,includingassaydesign.


TheMessageAmpPremierRNAAmplificationKitisthelatestinnovationforthepreparationofRNAsamplesformicroarrayanalysis.ThekitbuildsuponthelinearRNAamplificationmethodandreagentsdevelopedforMessageAmpIIKitswithenhancementsthatcreateasimplifiedworkflow,improvedperformance,andmoreflexibleRNAinputrequirements.AllMessageAmpkitsemploytheprovenmethodologyforRNAsamplepreparationandlabelingbasedontheoriginalT7invitrotranscription(IVT)amplificationtechnology,knownastheEberwineorreversetranscription-IVT(RT-IVT)method.TheRT-IVTmethodisconsideredthegoldstandardforsamplepreparationinmicroarray-basedexpressionprofiling.ItiswelldocumentedinthecurrentscientificliteratureandwasexperimentallyvalidatedusingTaqManRT-PCR.MessageAmpPremierKitfeatures:

LowestRNAinputrequirementsforasingleroundofamplification:20ngtotalRNAyieldsenoughbiotin-modifiedaRNAformostmicroarrayformats,withresultsequivalenttothosefromsamplespreparedfromsignificantlymorestartingmaterialusingotherRT-IVTkits
VeryflexibleRNAinputrequirements:20500ngtotalRNA,dependingonthetissueorcelltype
Streamlinedworkflow:dependingontheRNAinputamounts,onlyasingledayofprocessingtimeisrequiredtocompleteamicroarrayexperiment
Simplifiedprotocolandstreamlinedreagents
Completekit:everythingneededtopreparebiotin-modifiedaRNAisincluded
RigorousqualitycontrolthatincludesGeneChipanalysis
T7MEGAscriptIVTdeliversupto50,000-foldamplification
Single-tubeformat
Easy-to-use,high-recovery,magnetic-beadaRNApurification

UsingtheMessageAmp&tradePremierRNAAmplificationKit
Basedonthisprovenenzymaticamplificationandlabelingstrategy,theMessageAmpPremierprocedurestartswithasimplecDNAsynthesisreactionusingArrayScriptreversetranscriptase.NocleanupstepisrequiredaftertheRT;cDNAisuseddirectlyinahighyieldIVTreactionusingMEGAscripttechnology.TheIVTisconfiguredtoincorporatethemodifiednucleotide,biotin-UTP,intotheaRNAsynthesized.(Inthisprotocol,theamplifiedRNAisreferredtoasaRNA;intheliterature,itisalsocommonlycalledcRNA.)Oncepurified,theaRNAissuitableforuseonmicroarraygeneexpressionanalysisplatformsdesignedforbiotin-modifiedantisenseRNAsamples.
Getsensitivityandspecificityinaneasy-to-useone-stepqRT-PCRreaction.ThePowerSYBRGreenRNA-to-CT1-Stepkithasbeenformulatedtoensuremaximumsensitivityandreliability.Thenovelformulationreducesfalsepositiveresultscausedbyprimerdimers,significantlyimprovesdataaccuracy,andprovidesreliabledetectionoflowabundancetargets.Itincludes:
AmpliTaqGoldDNAPolymeraseUPforhotstartqPCR,minimizingnon-specificproductformation
ArrayScriptUPReverseTranscriptase,anengineeredRTthatproduceshighyieldsoffull-lengthcDNA
Anadditivethatreducesprimerdimerformation,reducingnon-specificproductsthatcancausefalsepositivesignals
RNaseinhibitor,topreventdegradationofRNAtemplates
AproprietaryROXdyethatservesasapassiveinternalreferencetonormalizenon-PCR-relatedfluorescencefluctuationsforsuperbprecisiononAppliedBiosystemsreal-timePCRinstruments.
ThisAmbionaRNAamplificationkithasextensiveimprovementsoverexistingmethods.Ithasfewercomponents,pipettingsteps,andmanipulationsintheworkflowwhichshortenhands-ontime,andreduceoperator-dependanterrorsandvariability.Thekitusesaglassfiber,filter-basedaRNApurificationmethod.Thekitincludessufficientreagentsfor10reactions.

SimplifiedWorkflow
Allreactionsoccurinasingletube
MasterMixformulationsreducepipettingstepsandhands-ontime
Shortenedprocessingtimeduetooptimizedincubationsandcondensedprocedure
Lesshands-ontimemeanslessopportunitytointroduceerrors

ImprovedReactionEfficiency
Startwithaslittleas20ngtotalRNAandproduceenoughaRNAforGeneChipanalysis
Startwith100ngoftotalRNA(witha4hourIVT)andhybridizetoaGeneChiparrayinasingleday

DNAmicroarraysareusedtoevaluatetheexpressionlevelsofthousandsofgenesinasingleexperiment.High-densityarraysarethemethodofchoiceforassessinggeneexpressiononagenome-widescale.Oneofthechallengesassociatedwithmicroarraytechnology,however,isthatadequateamountsoflabeledantisenseRNA(knownascRNAoraRNA)arerequired.T7linearamplification,commonlyreferredtoastheEberwinemethod,overcomesthischallengebysimultaneouslyamplifyingandlabelingtheRNA.ThismethodisthebasisoftheMessageAmpIIIRNAAmplificationKit.

WorkflowImprovements
Thekitincludesasetofmastermixes,reducingthenumberofcomponents,pipettingsteps,andmanipulations.StepssuchasinitialprimerannealingandcDNApurificationhavebeeneliminated.Allstepsofthebiotinlabeling/amplificationprocesstakeplaceinasinglereactiontube.Thissimplifiedworkflowsignificantlyshortenshands-ontime,reducingoperator-dependanterrorsandvariability.

ImprovedReactionEfficiency
TheMessageAmpPremierKitallowsaresearchertoperformexpressionprofilingwithaslittleas20ngofinputRNA.Thekitrequiresaslittleas100ngoftotalRNA,producingenoughlabeledaRNAtohybridizetoaGeneChiparrayinasingleday.RepresentativeyieldsfromtheMessageAmpPremierKitusingHeLatotalRNAandRNAfrommultipletissuetypesareshowninthedata.
TheHighCapacityRNA-to-cDNAKitisastreamlinedreversetranscriptionkitdesignedforoptimumperformancewithTaqManGeneExpressionMasterMix,PowerSYBRGreenPCRMasterMix,andotherPCRenzymes.

Featuresofthiskitinclude:
Fastshortreactiontime(typically3060min)
Convenienteasyworkflowwithfewpipettingsteps
Reliablereversetranscriptionofbothabundantandlimitedtargets
Flexibleoptimized2-stepprotocol,enablingmultiplePCRreactionsfromasinglereversetranscriptionreaction

StreamlineyourcDNAsynthesiswhilemaintainingdetectionsensitivity
Thecomponentsofthetwo-tubekitworktogethertoprovidesensitiveandspecificreversetranscriptionacrossabroadrangeoftemplateamounts.AmplificationofadilutionseriesoftheRNAconcentrationreferencestandarddemonstratesexceptionalreversetranscriptionandPCRefficiencyacross11ordersofmagnitude.Thiswidedynamicrangeallowsonesetofreactionconditionstodetecttranscriptsfromhighlyactiveaswellasweaklyexpressedgenes.Thetwo-tubeformulationreducesreactiontimeandrequiresfewerpipettingstepsthanwiththeHighCapacitycDNAReverseTranscriptionKit,whilemaintaininghighperformance.


AppliedBiosystemsTaqManMicroRNA反转录试剂盒提供最佳性能的基于TaqManMicroRNAAssay反应所需的所有成分。该试剂盒包含200个反应，试剂盒中的成分可结合TaqManMicroRNAAssay附带的反转录引物，从而将miRNA转换为cDNA。

这是一种高度特异性的试剂盒，可保证仅对成熟miRNA进行逆转录，不对miRNA前体反应。作为一种敏感的试剂盒，其可对有限的样品进行反应，仅需1-10ng的总RNA。适用于快速、简单且可扩展的2步法定量RT-PCR定量分析，从而在不到三小时内提供优质的分析结果。
TheBovineVirusDiarrheaRNATestKit(VetMAX-GoldBVDVPIDetectionKit)isintendedforuseintherapid,invitrodetectionofbovinevirusdiarrheavirus(BVDVRNA)extractedfromearnotchesinacuteandpersistentlyinfected(PI)cattle.

TheVetMAX-GoldBVDVPIDetectionKitisthefirstUSDA-licensedreal-timePCRtestforthedetectionofBVDVincattle.Theassayisasingle-tube,real-time,reversetranscription-polymerasechainreaction(RT-PCR)inwhichRNAisreversetranscribedintocDNA,andBVDVtargetsareamplifiedanddetectedinrealtimeusingfluorescentTaqManprobes.

HaveConfidenceinYourResults:
Provenrepeatabilityandreproducibilityinfieldstudies
Highlyspecificandsensitivedetectionoftype1andtype2BVDV
ConsistentlydetectsLimitations:
Thekitisnotintendedfordiscriminatingviralsubtypes
SamplesshouldbehandledasrecommendedtopreventdegradationofanyBVDVRNAthatispresent
RNAextractionmethodsshouldyieldRNAfreeofRT-PCRinhibitors,whichcanpreventamplificationoftargetRNA
FollowgoodPCRpracticestopreventfalsepositiveamplificationsduetocontaminationoftestsampleswithPCRproducts

Samplesize0.5-1mlovernightculture;Typicalyieldupto5g

Providescentrifuge-freepurification,idealforautomation
Avoidsintroductionofenzymaticcontaminantsforbetterresults
Optimizedforautomation

TheChargeSwitchNoSpinKitsprovidecentrifuge-freemicroprepplasmidpurification,idealforautomation.ThenovelNoSpinreagentcausesbacterialcellflocculationandallowscellstobepelletedwithouttheneedforautomationbottleneckssuchascentrifugation.Thiskitincludesallthenecessaryreagentsforthenumberofprepslistedandrequirestheuseofamagneticaccessory.PleaseseetheAccessoriesChapter(Chapter6)toselectthebestmagneticaccessoryforyourneeds.
ThePorcineIL-1BSimplexProcartaPlexkitmeasuresIL-1BproteinandisdesignedtobecombinablewithotherSimplexkitssothatyoucancreateyourownmultiplexpanelthatutilizesLuminexxMAPtechnologyforproteindetection/quantitation.WhencombiningmultipleSimplexkits(i.e.,whenyouarenotusingapre-configuredMultiplexPanel),onlyonebufferkit(soldseparately)isneededforeachassayplateregardlessofplexsize.

ProcartaPleximmunoassaysarebasedontheprinciplesofasandwichELISA,usingtwohighlyspecificantibodiesbindingtodifferentepitopesofoneproteintoquantitateupto80proteintargetssimultaneouslywhenusingtheFLEXMAP3DorLuminex200instrumentandupto50proteintargetswhenusingtheMAGPIXinstrument.ProcartaPlexassaysrequireaslittleas25Lofplasmaorserum,or50Lofcellculturesupernatant,andjustfourhourstoobtainanalyzedresults.

FlexiblepanelsdesignyourownpanelswithSimplexkitstomeasureyourownarrayoftargets
Moreresultspersamplemeasureupto80proteintargetsinasingle2550Lsample
Well-establishedLuminextechnologythemostreferencedmultiplexingplatformforproteindetectionandquantitation

TheLuminexMagPlexsuperparamagneticmicrospherebeadsintheProcartaPlexassayareinternallydyedwithpreciseproportionsofredandinfraredfluorophorestocreate100spectrallyuniquesignaturesthatcanbeidentifiedbytheLuminexxMAPdetec¬tionsystems(Luminex200,FLEXMAP3D,andMAGPIXsystems).SimilartoasandwichELISA,theProcartaPlexassayusesmatchedantibodypairstoidentifytheproteinofinterest.InaProcartaPlexmultiplexassay,eachspectrallyuniquebeadislabeledwithantibodiesspecificforasingletargetprotein,andboundproteinsareidentifiedwithbiotinylatedantibodiesandstreptavidinR-phycoerythrin(RPE).Theconjugationofprotein-specificantibodiestoadistinctbeadallowsforanalysisofmultipletargetsinasinglewell.

ThemostsignificantdifferencebetweenaProcartaPlexassayandELISAisthatthecaptureantibodyintheProcartaPlexassayisconjugatedtoamagneticbeadandnotadsorbedtothemicroplatewell,sotheProcartaPlexassayreagentsarefree-floatinginthesolution.Fordetection,theLuminex200instrument,forexample,containstwolasers,onetodistinguishthespectralsignatureofeachbeadandthesecondtoquantifytheamountofRPEfluorescence,whichisproportionaltotheamountofproteinpresentinthesample.ProcartaPlexmultiplexassayscanprofileupto80timesmoretargetproteinsusingsignificantlylesssampleinthesametimethatittakestoperformatraditionalsandwichELISA.

ProcartaPlexSimplexkitsprovidetheabilitytocreateyourownuniquepanel.Morethan90%ofProcartaPlexSimplextargetscanbecombined,providingyouwithsuperiorflexibilitywhencreatingyourownmultiplexpanel.

ProcartaPlexSimplexkitsareavailableacrosssixspecies(human,mouse,rat,nonhumanprimate,porcine,andcanine).Visitthermofisher.com/procartaplexformoreinformation,includingacomprehensivelistofindividualproteintargets.

Reactivity/Species
Pig

SuitableSampleTypes
cellculturesupernatant,serum,plasma

SampleVolume
Serum,Plasma:25L;CCS:50L

ReportedApplication
MultiplexImmunoassay


TheIonAmpliSeqMouseBCRIGHSRAssay,RNA,isarobust,targetednext-generationsequencing(NGS)assayforuseinbasicandtranslationalimmunology,immuno-oncology,hematooncology,andvaccineresearch.ItisdesignedtoaccuratelyidentifyandmeasuretheclonalexpansionofBlymphocytesinblood,peripheralbloodleukocytes(PBLs),peripheralbloodmononuclearcells(PBMCs),andfresh-frozen(FF)andformalin-fixedparaffin-embedded(FFPE)samples.TheassayidentifiesuniqueB-cellclonesthroughtargetingofthehighlydiversecomplementarity-determiningregion3(CDR3)oftheB-cellreceptor(BCR)immunoglobulinheavy(IGH)chainfromgenomicDNAtemplate.ThenucleotidesequenceoftheIGHCDR3regionservesasanaturalbarcodetoenableclonetrackingandmeasurementsofB-cellclonalexpansionanddiversity.AnalysisofIGHCDR3-regionaminoacidmotifsmayrevealsignaturesofB-cellresponsestodefinedantigens.

BenefitsoftheIonAmpliSeqMouseBCRIGHSRAssay,RNA,include:
Compatibilitywithavastarrayofresearchsampletypes,includingFFPEtissue,fresh-frozen(FF)tissue,wholeblood,PBLs,andPBMCs
Highsensitivityandlowlimitofdetection(LoD)forrarecloneidentificationthroughdual-barcodeindexing
Efficientworkflowwith48hoursample-to-resultstime
Flexibleinputrequirementsrangingfrom100ngto1g
Streamlinedanduser-friendlyinformaticssolutionwithwithmulti-sampleanalysisfunctionality

TheIonAmpliSeqMouseBCRIGHSRAssay,RNA,providesasinglepoolofmultiplexPCRprimersandlibraryreagentstogenerate90-bpampliconsthatcanbesequencedonallchiptypessupportedbytheIonGeneStudioS5sequencingsystems,allowingyoutopickthebestmultiplexingconfigurationforyouruniquesamplebatchingneedsandthroughputrequirements.Theentireworkflow,(fromlibrarypreparationtoanalysisofsamples,canbeaccomplishedintwodaysusingtheIonCheftemplatingsystemandIonGeneStudioS5system..

TheIonAmpliSeqMouseBCRIGHSRAssay,RNA,withitshighsensitivityandspecificity,supportskeyapplicationsinimmunology,immuno-oncology,hemato-oncology,andvaccineresearch.Thehigh-sensitivitydual-barcodeindexing,flexibleinputrequirements(100ng1g),high-depthsequencing,andhigh-throughputcapabilitymakethisassayidealfortestingvarietyofhypothesisforbasicortranslationalbiomarkerresearch.Asmallfragment-sizerequirementfortheinputmaterialsforlibrarycreationandhighmultiplexingontheIon530ChipoftheIonGeneStudioS5sequencerenablesresearcherstofocusontestingavarietyofusecasesinmousemodels,includingtheroleofTcellsingeneratingimmuneresponsetoimmunotherapiessuchascheckpointblockadeinhibitors,cancervaccines,orchimericantigenreceptor(CAR)T-celltherapies.

Note:InformationaboutdataanalysisusingIonReporterSoftwarev5.12canbefoundintheUserGuidebelow.


TOPOTACloningkitsforsequencingprovideahighlyefficient,5-minutecloningreactionforthedirectinsertionofTaqpolymerase-amplifiedPCRproductsintoaplasmidvectorforsequencing.EachkitusesthepCR4-TOPOTAvectorwithspeciallydesignedsequencingprimersitesthatreturnmoreinsertsequenceandlessvectorsequencefromeachreaction,andareavailablewithavarietyofcompetentcells,ornocompetentcells,dependingonyourneedsandbudget.ThesekitsincludethetoolsnecessarytocloneandselectrecombinantvectorscontainingyourPCRfragmentofchoice.

Getmoresequenceallowsformoreinsertsequenceandlessvectorsequencewhenusingstandardsequencingprimers
FastandeasygofromPCRtoclonesinjust3steps,includinga5-minutecloningreaction
Efficientobtainupto95%cloneswithcorrectinsert
Provenenablingreliableperformanceforoveradecadewithover20,000citations

pCR4-TOPOTAVectorOptimizedforSequencing
ThepCR4-TOPOTAvectorhasaminimizedmultiplecloningsiteand,thus,ashorterdistancebetweenthesequencingprimersitesandtheinsertsite(aslittleas33bp).Sosequencingreactionsproducelessvectorsequenceandmoreinsertsequence.ThepCR4-TOPOTAvectorhassitesfor4commonsequencingprimers:M13forward,M13reverse,T7,andT3.Thekitsincludeanaliquotofeach.

pCR4-TOPOTACloneSelectionandManipulation
ThepCR4-TOPOTAvectorcontainsbothampicillinandkanamycinresistancemarkersandaLacZ-ccdBgenefusionforpositiveselectionandblue/whitescreening.ThevectorsminimizedmultiplecloningsitestillincludesflankingEcoRIsitesforsimplifiedexcisionofclonedPCRproductsandauniqueSse8387Isiteforgenerationofnesteddeletionspriortosequencing.T7andT3promotersarealsopresentforinvitrotranscription.

SimplifiedTOPO-basedCloning
UsingTOPOcloningtechnology,thereisnoneedforPCRprimerscontainingspecificsequences,post-PCRprocedures,vectorpreparation,orothertime-intensiveDNAmanipulationsteps.JustaddyourPCRreactionstraighttotheprovidedtopoisomerase-chargedvector,incubate5minutes,andtransformE.colicompetentcells.

EfficientCloning
Withupto95%ofclonescarryingthedesiredinsert,youcanscreenlessclonesandsavetimeandmoney.ThepCR4-TOPOTAvectorusedinthiskitcomeswith3ToverhangsforefficientligationofTaq-amplifiedPCRproducts,whichcontain3Aoverhangs.

TheMostWidelyUsedCloningKit
Whenitcomestocloning,TOPOcloningtechnologyhasbeenareliablepartnerforthousandsofscientistsforovertenyears.Fast,simple-to-use,andefficient,TOPOcloninghasbeenappliedtomanydifferentvectorsforawidearrayofapplications.

KitOptions:TOPOTACloningKitsforSequencing
TOPOTACloningkitsforsequencingcanbepurchasedwithavarietyofcompetentcellsthatdeliverdifferentadvantagesdependinguponyourneeds:
Generalcloning:TOP10(Cat.Nos.K4575-J10,K4575-01,K4575-40)
High-efficiencycloning:TOP10Electrocompcells(Cat.Nos.K4580-01,K4580-40)
Generalcloning,bacteriophageT1resistance:DH5-T1R(Cat.Nos.K4595-01,K4595-40)
Fastgrowth:Mach1-T1RchemicallycompetentE.coli(Cat.No.K4530-20)
Provideyourowncells(Cat.Nos.450071,450030)

WealsoofferaversionofthekitthatincludesaPureLinkQuickPlasmidMiniprepKit(Cat.No.K4575-02)foruseinisolationofclean,sequencing-ready,recombinantplasmid.
TheVybrantDyeCycleViolet/SYTOXAADvancedapoptosiskitisusedfortheidentificationofapoptoticcells.Thiskitprovidesarapidandconvenientassaybaseduponfluorescenceanalysisofthecompactedstateofthenuclearchromatininapoptoticcells.Thekitcontainssolutionsofthecell-permeantVybrantDyeCycleVioletstain(Ex/Emmax~370/440nm)whichisexcitedwithUVorvioletlaserandSYTOXAADvanced(Ex/Emmax~546/650nm)whichisexcitedwitha488nmlaser.SYTOXAADvancedisusedtodetectchangesinpermeabilityoftheplasmamembranethereforeallowingidentificationofdeadcells.Thestainingpatternresultingfromsimultaneoususeofbothdyesincombinationmakeitpossibletodistinguishnormal,apoptotic,anddeadcellpopulationsbyflowcytometry.

Viewaselectionguideforallapoptosisassaysforflowcytometry.
OurnewPro-QDiamondphosphoproteingelstainprovidesaconvenientmethodforselectivelystainingphosphoproteinsinacrylamidegels,withouttheneedforblottingorphosphoproteinspecificantibodiesandWesternblotanalysis.Thisreagent,whencombinedwithourtotalproteinstainSYPRORubyproteingelstainbecomesapowerfulcombinationforquantitativeproteomeanalysis.ThisconvenientPro-QDiamondphosphoproteingelstainandSYPRORubyproteingelstainpackprovideseachstainin1literamounts.Thiscombinationisalsoavailablein200mLamounts(M-33306).
TheIonTorrentOncomineBCRIGHLRAssayisarobust,targetednext-generationsequencing(NGS)assaydesignedforuseinhematology-oncology,immuno-oncology,infectiousdisease,andbasicimmunologyresearch.Unlikeothertechnologies,theOncomineBCRIGH-LRAssayoffersmorethan400baseampliconsoflibrarysequencingthroughlong-readsequencingchemistry,enablingcomprehensivecoverageoftheB-cellreceptor(BCR)immunoglobinheavy(IGH)chain.TheassaykitincludesasinglepoolofmultiplexPCRprimers,cDNAsynthesiskit,andIonAmpliseqlibraryreagents.

TheOncomineBCRIGHLRAssayisdesignedtoaccuratelymeasureclonalexpansion,quantifysomatichypermutation,andisotypeBlymphocytesusingtotalRNAextractedfrombonemarrow,wholeblood,peripheralbloodleukocytes(PBLs),peripheralbloodmononuclearcells(PBMCs),orsortedcellsinfresh-frozenresearchspecimens.TheassayutilizesIonAmpliSeqmultiplexPCRtechnology.Itamplifiesframework1(FR1)andtheconstantgeneregionoftheBCRIGHgenetointerrogatehighlydiversecomplementarity-determiningregionsCDR1,CDR2,andCDR3,aswellastheCH1domainoftheconstantgenetoenabledistinctionofallnineisotypes.IGHCDR-regionaminoacidmotifsmayrevealsignaturesofB-cellresponsestodefinedantigensforuseasfuturemarkersofautoimmuneorinfectiousdisease.Automatedclonallineageanalysisandmulti-sampleanalysiscapabilityfacilitatestrackingofB-cellclonalevolutionarisingfromsomatichypermutationandclassswitchrecombinationinresearchsamples.

OncomineBCRIGHLRAssaybenefitsinclude:
•Extendedampliconallowsforquantificationofvariablegenesomatichypermutationandisotypeidentificationtoenablehemato-oncology,immuno-oncology,andinfectiousdiseasetranslationalandclinicalresearch
•Assayprimersaredesignedtoamplifyallvariableandjoininggeneallelesinthegold-standardIMGTdatabase
•RNAinputimprovesdetectionofchangesinplasmablastandplasmacellpopulationsinresearchsamplesfollowingimmunechallengeoradministrationofimmunomodulatoryagents
•SuperiormultiplexPCRdesignthroughIonAmpliSeqtechnologyassureshighaccuracyandhighsensitivity
•Ultra-lowsubstitutionerrorrateminimizesartifactsarisingfromsequencingerrorstoenablehighlyaccurateassessmentofsomatichypermutationandclonalheterogeneityinmalignantB-cellresearchsamplesofinterest
•TheentireworkflowfromisolationofRNAtoanalysisofsamplescanbeaccomplishedwithintwodaysusingtheIonCheftemplatingsystemandIonGenestudioS5sequencingsystem
•Flexibleinputrequirementsensuresuccessfullibraryconstructionwithaslowas25ngandupto2gtotalRNAinput
•Streamlinedanduser-friendlyinformaticssolutionoffersautomatedclonotyping,somatichypermutationquantification,clonallineageanalysis,reportingofkeyrepertoirefeatures,andmulti-sampleanalysiscapabilitytotrackB-cellclonesacrossresearchsamples

Learnmoreabouttheassay

Note:InformationaboutdataanalysisusingIonReporterSoftwarev5.12canbefoundintheUserGuidebelow.

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