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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
    AgeI11
    AhdI11
    AlwNI21
    ApaI11
    ApoI11
    AseI0.31
    AvaI101
    AvrII11
    BaeI31
    BamHI31
    BanII11
    BglII81
    BpmI11
    BsaAI201
    BsaI21
    BsaWI31
    BsaXI21
    BsgI11
    BsmI11
    BspDI11
    BspEI11
    BspMI**1
    BspQI31
    BsrFI21
    BsrGI11
    BssHII41
    BtgI51
    ClaI51
    EagI101
    EcoNI31
    EcoO109I81
    EcoRI31
    EcoRV11
    HincII41
    HindIII51
    KasI41
    KpnI21
    MluI21
    NarI101
    NcoI11
    NdeI31
    NheI51
    NmeAIII**1
    NruI11
    NsiI11
    PciI31
    PsiI31
    PstI11
    PvuI21
    PvuII21
    SacI51
    SalI101
    SapI11
    ScaI201
    SmaI11
    SnaBI11
    SpeI11
    SphI31
    SspI41
    StuI31
    StyI41
    TliI21
    TspMI11
    Tth111I21
    XbaI21
    XhoI101
    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.