SP6 In Vitro Transcription Reaction Protocol (M0207) - NEB

SP6 In Vitro Transcription Reaction Protocol (M0207) ... For analysis of transcription run 1 µl of transcription reaction on a 1% agarose gel. Note: Higher yields ... Home Protocols SP6InVitroTranscriptionReactionProtocol(M0207) SP6InVitroTranscriptionReactionProtocol(M0207) Setupthefollowingreactiononice (Pleasenotethatenzymeshouldalwaysbeaddedlast): COMPONENTS 20µlREACTION FINALCONC.ORAMOUNT RNAPolReactionBuffer 2µl 1X NTP Xµl 0.5mMeach H2O Xµl   TemplateDNA Xµl 1µg RNaseInhibitor(optional) 0.5µl 1U/µlfinal FreshDTT(optional) Xµl 5mMfinal SP6RNAPolymerase 2µl   Gentlymixthereactionbypipettingupanddownandmicrofugebriefly. Incubateat37°Cfor1hour.Forshortertranscripts,incubateat37°Cfor2-16hours. Foranalysisoftranscriptionrun1µloftranscriptionreactionona1%agarosegel. Note:HigheryieldsofRNAmaybeobtainedbyraisingrNTPconcentrationsupto4mMeach.MgCl2concentrationsshouldalsobeincreasedupto20mM(4mMabovethetotalrNTPconcentrationof16mM).   Chooseyourcountry NorthAmerica Canada UnitedStates Europe France Germany UnitedKingdom Asia-Pacific Australia China Japan NewZealand Singapore Ifyoudon'tseeyourcountryabove,pleasevisitour internationalsite SessionExpired Youhavebeenidleformorethan20minutes,foryoursecurityyouhavebeenloggedout.Pleasesignbackintocontinueyoursession. SignIn InstitutionChanged YourprofilehasbeenmappedtoanInstitution,pleasesignbackforyourprofileupdatestobecompleted. SignIn SignintoyourNEBaccount Tosaveyourcartandviewpreviousorders,signintoyourNEBaccount.Addingproductstoyourcartwithoutbeingsignedinwillresultinalossofyourcartwhenyoudosigninorleavethesite. SignIn SignIn ContinueasGuest Don'tshowmeagain