Description:
CompleteKitKAPARNAHyperPrepKitswithRiboErase(HMR)
Single-DayRNA
TheKAPARNAHyperPrepKitswithRiboErase(HMR)utilizenovelchemistrythatenablesthecombinationofenzymaticstepsandfewerreactionpurifications,resultinginatrulystreamlinedsolutionforthepreparationofhigh-qualityrRNA-depletedRNA-seqlibraries.Byutilizingatargetedenzymaticmethodfordepletion,ourworkflowenablesefficientreductionofribosomalRNA(rRNA)andamorecompleterepresentationofthetranscriptome,includingprecursormRNAsandnon-codingRNA(ncRNA).Thestrand-specificworkflowisflexIBLe–supportinglibraryconstructionfromlower-inputamountsanddegradedsamples.KitscontainallreagentsrequiredforrRNAdepletionandlibrarypreparation,withtheexceptionofKAPAAdapters(availableseparately).Benefitsinclude:
- single-daylibraryconstruction,inclusiveofrRNAdepletion
- reducedhands-onandoveralltimethroughfewerenzymaticandreactioncleanups
- flexibleinputof25ng–1µgtotalRNAfromhuman,mouse,orratspecies*
- highersuccessrateswithlowerinputanddegradedsamples
- maintainover99%strandspecificity*
- KAPAPureBeadsincludedforreactionpurifications
NEW!KAPADual-IndexedAdapterKitsarenowavailable. FormoreinformationonKAPAAdapterKits,scrolldowntotheOrderingsection,ordownloadtheKAPAAdapterandBeadCalculator.DownloadourAdapterandBeadCalculator
*Dataonfile.
ForResearchUseOnly.Notforuseindiagnosticprocedures.

ProductHighlights
Sequencewhatmatters
- WastefewerreadsduetothecombinationofrRNAcarryoverandPCRduplicates
- Identifymoreuniquetranscriptsandgeneswithequivalentsequencing
Achievesuperiorcoverageuniformity
- Obtainmoreuniformdistributionofreadsacrosstranscripts
- ImprovecoverageofdifficultGC-richtranscripts
Generatehigh-qualitylibrariesfromdegradedsamples
- Inputaslittleas25ngwithFFPEsamples,dependingontotalRNAquality
- Achievelowduplicationratesandhighlyefficient,reproduciblerRNAremovalwithdegradedsamples
- Identifymoreuniquetranscriptsandgeneswithequivalentsequencing
Achievereliableresultswithdegradedinputs
- Attainahigh-degreeofexpressioncorrelationbetweenpairedFFPEandfreshfrozensampleswhichprovidesincreasedconfidenceinsequencedataaccuracy
RelatedProducts
Areyousequencinglow-input,FFPEorhighqualityDNA? RNA? CheckouttheseKapaNGSproductstoimproveyourworkflowandresults:
KAPAmRNAHyperPrepKits
KAPAStrandedmRNA-SeqKits
KAPALibraryAmplificationKits
KAPAHyperPrepKits
KAPALibraryQuantificationKits
Applications:
Applications- Wholetranscriptome
- Geneexpressionanalysis
- Detectionofgenefusions,isoforms,andotherstructuralvariants
- Noveltranscriptidentification,includingnoncodingtranscripts
- SNVdiscovery
KitSpecificationsandContents/Storage:
KitSpecificationsandContents/StorageEnzymesandbuffersforrRNAdepletion,CDNAsynthesis,andlibrarypreparationcanbestoredforupto10monthsat-20°C.KAPAPureBeadscanbestoredforupto10monthsat4°C.(USonly)
KitscontainHybridizationBuffer,HybridizationOligos(HMR),DepletionBuffer,RNaseH,DNaseBuffer,DNase,Fragment,PrimeandEluteBuffer(2X),1stStrandSynthesisBuffer,KAPAScript,2ndMarkingBuffer,2ndStrandSynthesis&A-TailingEnzymeMix,LigationBuffer,DNALigase,PEG/NaClSolution,KAPAPureBeads,LibraryAmplificationPrimerMix(10X),andKAPAHiFiHotStartReadyMix(2X).

Specifications
- SpecDescription
- CompatiblePlatform0616
- LibraryTypeRNA
- StartingMaterialHigh-qualityanddegradedtotalRNA
- InputAmount25ng–1µg
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植物材料 用最精确的方法,称取不超过100mg的植物材料,置于处理过的研钵,加入液氮进行研磨 将研磨得到的粉末,快速转移至无RNase,并经过液氮冷却的2mL离心管(自备),
各种tRNA的一级结构互不相同,但它们的二级结构都呈三叶草形。这种三叶草形结构的主要特征是,含有四个螺旋区、三个环和一个附加叉。四个螺旋区构成四个臂,其中含有3′末端的螺旋区称为氨基酸臂,因为此臂的3′-末端都是C-C-A-OH序列,可与氨基酸连接。三个环分别用Ⅰ、Ⅱ、Ⅲ表示。环Ⅰ含有5,6二氢尿嘧啶,称为二氢尿嘧啶环(DHU环)。环Ⅱ顶端含有由三个碱基组成的反密码子,称为反密码环;反密码子可识别mRNA分子上的密码子,在蛋白质生物合成中起重要的翻译作用。环Ⅲ含有胸苷(T)、假尿苷(ψ)、胞苷(C),称为TψC环;此环可能与结合核糖体有关。tRNA在二级结构的基础上进一步折叠成为倒“L”字母形的三级结构(图3-2-6)。
tRNA分子中稀有碱基的数量是所有核酸分子中比例最高的,这些稀有碱基的来源是转录之后经过加工修饰形成的。
tRNA共有61种,对应61种氨基酸的密码子(64种密码子中,2个是起始密码子,且对应一定氨基酸;3个是终止密码子,不对应氨基酸)
1、测浓度和纯度,是否达标
2、跑电泳看条带是否与测的数据相互印证
如果上述两点满足,就不需要纯化,否则需要进一步纯化。
不过根据经验,一般都不需要,我们实验室用的是GNT系列的试剂盒,提取结果就直接进入下游实验了,效果挺稳定
质粒是真核细胞细胞核外或原核生物拟核区外能够进行自主复制的遗传单位,包括真核生物的细胞器(主要指线粒体和叶绿体)中和细菌细胞拟核区以外的环状脱氧核糖核酸(DNA)分子.现在习惯上用来专指细菌(大肠杆菌)、酵母菌和放线菌等生物中细胞核或拟核中的DNA以外的DNA分子.
小鼠没有叶绿体,你可以直接参照小鼠线粒体的提取方法进行提取.
实验非常不顺,想构建CDNA文库,但是从mRNA开始屡次失败,考虑主要是纯化过程中损失过多。想从总RNA入手,但是不知道实验步骤。不知那位大侠能提供总RNA建库的实验步骤。另外我现在手头有OligoDT,RT酶,苦于没有第二连合成试剂盒,不知道能否用普通PCR试剂盒Teq酶替代第二链合成过程中的DNA聚合酶I。好像有种方法合成第二链时,不需要另外的引物,利用降解的RNA作引物即可,我想直接设定PCR两个循环,合成第二链,不知方法可行否?愁啊,等着毕业,时间紧急,恳请帮忙。谢谢。










