·RNAGel(CrawfordLab)

·GelElectrophoresisofRNA(BeverlyFaulkner-Jones)greattipsonRNAgelelectrophoresis.

·NorthernGelandTransfer(gopher://iubio.bio.indiana.edu)UsingglyoxalatedRNA

·RNAElectrophoresis(MolecularGeneticsRecLab)

·RNAElectrophoresisinDenaturingFormaldehydeGel(MolecularGeneticsRecLab)

·Formaldehyde-AgaroseGels(BeverlyFaulkner-Jones)

Agarosegelshaveasmallerresolvingpowerthanpolyacrylamidegelsbutagreaterrangeofseparation-from200bpto>50kbusingstandardgelsandelectrophoresisequipment.RNAsupto10000kbcanbeseparatedinagarosegelsusingpulsedfieldgelelectrophoresis.

·DenaturingpolyacrylamidegelelectrophoresisofRNA(BeverlyFaulkner-Jones)Polyacrylamidegelshaveenoughresolvingpowertoseparatefragmentsdifferingbyonlyonebasepairinsize,buttheirrangeis~5to1000bp.Theyaremuchmoredifficulttohandlethanagarosegels.

·RecoveryofRNAfragmentsfrompolyacrylamidegels(BeverlyFaulkner-Jones)Thebestmethodisthecrushandsoakmethod.Itcanbesuedtoisolatesingle-strandedRNAfromdenaturinggels.TheRNAisofhighpuritybutthemethodislongandinefficientiflongtranscriptsaretoberecovered(cf.lessthan30%ofDNAgreaterthan3kbisrecovered).

  • PolyacrylamideGelElectrophoresis(PAGE)ofRNA(FrankBottone)ForusewithRibonucleaseProtectionAssay(RPA):

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