Description:
ThestudyofgeneexpressionoftenneedsRNApreparationfollowedbyCDNAsynthesisandPCR,butmostofthetime,youdon"twanttowastealargeamountofcellsforRNApreparation.Thisisaparticularproblemforresearchersusinglaserdissectedsamples,FACSsortedcells,andculturedcellsin96-wells. Signosis’DirectcDNA celllysisbufferallowsresearcherstopreparecelllysatefromsmallsamples,whichcanbeusedfordirectreversetranscriptionwithoutRNApreparation.CelllysatemadefromevenafewcellsisgoodenoughforreversetranscriptionofRNAtocDNA,whichyoucanthenuseforPCRanalysisorcDNAplatearrayassays.Principle:
Thebuffersareoptimizedforcelllysatepreparationandreal-timeRT-PCR.
Data:
ComparisonofPCRproductsamplifiedfromlysatesmadewithSignosisDirectcDNAcelllysisbufferorTRIzolwithoutDNAsetreatment. Byusingintron-spanningprimers,DNAamplifiedfromgenomicDNAandreverse-transcribedmRNAcanbedistinguished. PCRamplificationfromRNApreparedwithTRIzolcontainsabandthatcorrespondswiththepredictedlengthofthegenomicDNA,aswellasthepredictedlengthofthemRNA.However,PCRamplificationfromcell lysatepreparedwithSignosisDirectcDNAcelllysisbufferonlycontainsabandthatcorrespondswiththepredictedlengthofthemRNAandnotthegenomicDNAcontamination. |
