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《限制性内切酶酶切的常见问题及解决方法（common problems and solutions of restriction endonuclease digestion）》由会员上传分享，免费在线阅读，更多相关内容在教育资源-天天文库。
1、限制性内切酶酶切的常见问题及解决方法（Commonproblemsandsolutionsofrestrictionendonucleasedigestion）ThecommonproblemsandsolutionsofrestrictionendonucleasedigestionRestrictionenzymedigestionofthecommonproblemsandsolutions,personallyfeelverywellsummedup,speciallypostedforyoutoshare.Enzymedige
2、stionproblems,seeenzymeinstructions,thecorrespondingreagentcompanydirectory.Theenzymesproducedbydifferentcompanies,thesourceofstrains,thepreparationprocess,purity,viabilityandenzymaticactivitymaybeoptimizeddifferently,.Canbefoundintheabovedefinition,unitofenzymepreservat
3、ionconditions,enzymesystemof[bufferandotherenzymessuchasbuffer,doublecutenzymereactiontemperature[someenzymesin50,55or30temperaturereaction,whethermethylationaffectstheenzymeactivityandwhetherthereisanasterisk,asteriskmayappearlivefactorsandprotectivebase,isocaudarner,is
4、oschizomer1.enzymescannotbecutorincompleteThe1.1plasmidisthemostcommonorpoorlyexpressedinhibitor.Thepresenceofmiscellaneousproteinscanaffectenzymedigestion,showingA260/A280below1.8;inhibitorsarecommoninphenols,salts,andethanol;[reDNA,usingreliablekitsorreliablemanualextr
5、actionreagents,1.2enzymeproblems:makesuretheenzymeiseffective[althoughmanyenzymeshaveexpiredtime,butexpiredaslongastheenzymecanbeeffectivelycut,available.Makesuretheenzymeisnoteffective.Markitandreplaceit.[note]topic:byendonucleaseA.endonuclease,ifnospecialrequirement,ke
6、ep-20~-30degrees.Isnotaslowaspossible,theenzymeisusuallypreservedin50%glycerolbuffer,whenthetemperatureistoolow,willfreeze(enzymetransportprocessexception,becausetheenzymerequireslowtemperaturetransport,sotheconvenientsituationistheuseofdryice,theenzymewillfreeze,butthef
7、reezingtimesislimited,isarelativelybetterchoice),ifisoftenused,theenzymewillberepeatedfreezingandthawing,therebyreducingtheactivity.Ofcourse,ifthetemperatureistoohigh,haha,youwanttogo.B.enzymeshouldbeusediniceboxforenzymeextraction.It'scleartoeveryone,butit'snoharmputtin
8、gtoomuchemphasisonit.Someofthestudentsbecausethelabisputout,theenzymeintheicebox,buttherearetwoignored:
