一种有效提取无核荔枝幼胚总RNA的方法.pdfVIP

维普资讯 AgriculturalScienceTechnology。2008。9(1)：29—31 Copyright◎2008。InformationInstituteofHAAS．Allrightsreserved AgriculturalBio-technology An Effective Method forExtracting TotalRNA from Young Embryo of SeedlessLitchi LIUXing．di一．ZHENGXue—qin 1．，瑚疵 eofTropicalBioseienceandBioteehnologyCATAS，Haikou57110l；2．AgriculturalCollegeofHainanUniversity，Danzhou571737 AbStract AtotalRNAextractionmethodforyoungembryoofseedlesslitchiwasintroduced．CTAB，Phenol(saturatedwithwater)，chloroform，Guani- dineisothiocyanatewereusedas mainextractionreagents．Polyphenoliecompoundswere removedeffectivelybyaddedPVPintotheextractionbuffersolu- tion．RNA was purifiedintensivelybyphenol，chloroofmr extraction，andethanoldepositionafterdepositedbyLiC1．Boththeresultsoffomr aldehydedena- turedagarosegeleleertophoresisandultravioletspcetrophotometeranalysisshowedhighintegitynadpuri~ofRNA．SothequalityofextractedRNAcould meetthedemandof mostmolecularbiologyexperimentsthatrequire higherqualityRNA． Keywords Seedlesslitehi：Youngemb ryo；RNA extraction RNAextractionisthebasicandkeyexperimentofmole． nylpyrrolidone)，2％ 3-mercaptoethanol(addedjustbeforeu— culebiologicalresearch．IfthequalityofRNA isnotgood． sing)．Denaturde solutionofgunaidinium thiocynaatewas theg0odexperimentresultwillnotbeobtainedorsomeprob— comprisedof4 mol／L guna idinium thiocynaate，25mmol／L lemsarestillexsitingforfurturestudies suchas rapidam- sodium citrate(pH=7)，0．5％ sodium lauroylsracosine ． plificationofcDNA ends，differential displayreversetran- Allglasswraeweredriedat180℃ overnightwhileplastics scription．PCR．northerncrossandinvitro trnaslation．Poly． weresokaedin0．1％DEPCfdiethylpyrocrabonate)at37℃ saccharide，polyphenol，protein and othersecondary metabo· for2ho