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  • Cleavage of Supercoiled DNA

    Restriction endonucleases cleave different DNA substrates with varying efficiency. Restriction enzymes were tested on their ability to cleave various plasmids (pBR322, pUC19 and pLITMUS) under standard reaction conditions. Single sites were tested on each of these plasmids, depending on availability, and average values were taken when there was more than one data point available.

    EnzymeUnits to Cleave
    PlasmidLambda
    AatII31
    Acc65I11
    AccI41
    AflII21
    AflIII11
    AgeI (1) (2)11
    AhdI11
    AlwNI21
    ApaI11
    ApoI11
    AseI0.31
    AvaI101
    AvrII11
    BaeI31
    BamHI (1)31
    BanII11
    BglII81
    BpmI11
    BsaAI201
    BsaI (1)21
    BsaWI31
    BsaXI21
    BsgI11
    BsmI11
    BspDI11
    BspEI11
    BspMI**1
    BspQI31
    BsrFI21
    BsrGI (1)11
    BssHII41
    BtgI51
    ClaI51
    EagI (1)101
    EcoNI31
    EcoO109I81
    EcoRI (1) (2)31
    EcoRV (1) (2)11
    HincII41
    HindIII (1)51
    KasI41
    KpnI (1) (2)21
    MluI (1)21
    NarI101
    NcoI (1) (2)11
    NdeI31
    NheI (1) (2)51
    NmeAIII**1
    NruI (1)11
    NsiI (1)11
    PciI31
    PsiI31
    PstI (1)11
    PvuI (1)21
    PvuII (1)21
    SacI (1)51
    SalI (1) (2)101
    SapI11
    ScaI (1) (2)201
    SmaI11
    SnaBI11
    SpeI (1) (2)11
    SphI (1)31
    SspI (1)41
    StuI31
    StyI (1)41
    TliI21
    TspMI11
    Tth111I21
    XbaI (2)21
    XhoI (2)101
    XmnI51
    ** BspMI and NmeAIII require 2 copies of its recognition sequence for cleavage to occur. Thus, the single BspMI site in pBR322 and pUC19 as well as the single NmeAIII site in pUC19 are resistant to cleavage. A 100-fold overdigestion with BspMI cuts less than half the DNA present, while a 10-fold overdigestion with NmeAIII cuts less than half the DNA present. Other plasmids may also be resistant to BspMI and NmeAIII cleavage.
    (1) A HF version of this enzyme is available.
    (2) A RE-Mix version of this enzyme is available.