
RapidextractionofRT-PCR-readyRNAfromculturedmammaliancells
Fast:Extractend-pointandreal-timeRT-PCRreadyRNAinminutesfromlowtohighnumbersofsamples
- Simple:Singletubeprotocol
- CompatIBLe:ExtractedRNAiscompatiblewithbothreal-timeandendpointRT-PCR
- Automation-friendly:Simpleprotocolmakesincorporationintoanautomatedworkfloweasy
- Flexible:ADnaseIdigestionstepcanbeaddedforsensitivedownstreamapplicationswhennecessary
- Safe:Notoxicreagentsused
TheQuickExtract™RNAExtractionKitisafast,simplewayofpreparingRNAforRT-PCR(bothend-pointandreal-time).Thesingle-tubesystemrequiresonlyvortexmixingtolysethecells,andpreparetheRNAforCDNAsynthesis.Theresultiseasyprocessingofonetohundredsofsamplesinminutes,withnosamplelossortoxicorganicsolvents.TheQuickExtractRNAExtractionSolutionworkswithculturedadherentandsUSPensioncellsincludingbuccalcells,andhasbeentestedonhuman,mouse,rat,E.coli,andS.aureuscellcultures.Itisnotsuitablefortissuesamplesorplantsamples.AnoptionalDNaseItreatmentmayimprovecertaindownstreamapplications.
Applications
- PreparationofRNAfromculturedadherentandsuspensioncellsforRT-PCR.
![]() | Figure1.End-pointRT-PCRofdifferentregionsofa14kbmessageusingHeLacellextractwiththeQuickExtract™RNAExtractionKit.Asamplecontaining105HeLacellswaslysedin100µLofQuickExtractRNAExtractionSolutionbyvortexmixing.ThelysatewasreversetranscribedwiththeMMLVReverseTranscriptase1stStrandcDNASynthesisKitusingstandardconditionsandrandomprimers.ThecDNAwasthenamplifiedwithsixprimersetstop532usingtheFailSafe™PCRSystem.LaneM,100 bpladder;Lane1,12984to13892;lane2,9406to10202;lane3,5194to5802;lane4,4191to4690;lane5,2280to2676;andlane6,1029to1329. |
![]() | Figure2.ComparativeyieldofRT-PCRproductwithdifferentRNAextractionkits.Lysateswerepreparedaccordingtomanufacturers'instructionsandusedastemplatetoproducecDNAusingtheMMLVRT1stStrandcDNASynthesisKit,followedbyPCRusingtheFailSafe™PCRSystemwithprimersfortheALDOAgene.Productswereseparatedona2%agarosegelandstainedwithSYBR®gold.LaneM,100bpladder;lanes1and2,QuickExtract™RNAExtractionKit;lanes3and4,kitfromVendor1;lanes5and6,kitfromVendor2. |
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(1)优级纯试剂 亦称保证试剂,为一级品,纯度高,杂质极少,主要用于精密分析和科学研究,常以GR表示。
(2)分析纯试剂 亦称分析试剂,为二级品,纯度略低于优级纯,杂质含量略高于优级纯,适用于重要分析和一般性研究工作,常以AR表示。
(3)化学纯试剂 为三级品,纯度较分析纯差,但高于实验试剂,适用于工厂、学校一般性的分析工作,常以CP表示。
(4)实验试剂 为四级品,纯度比化学纯差,但比工业品纯度高,主要用于一般化学实验,不能用于分析工作,常以 LR表示。
以上按试剂纯度的分类法已在我国通用。根据化学工业部颁布的“化学试剂包装及标志”的规定,化学试剂的不同等级分别用各种不同的颜色来标志,见表1。
表1 我国化学试剂的等级及标志
bhclna2so4nano3na2co3
chclnaohna2co3nacl
dba(oh)2nahco3alcl3nahso4
刚入这行,谢谢大家
A.H2SO4Na2SO4AgNO3BaCl2.
B.NaOHNa2CO3NaHSO4MgCl2
C.CaCl2NaNO3MgSO4BaCl2
D.HNO3KOHKClK2CO3

