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  • Restriction Endonucleases - Survival in a Reaction

    Restriction endonucleases vary with respect to their ability to maintain activity in a reaction over an extended period of time.

    While most routine restriction digests are incubated for one hour or less, there are certain applications that require the addition of less than 1 unit/µg of DNA and increasing the reaction time beyond one hour. The table below can be used as a guide when low levels of enzyme and extended reaction times are needed.

    For example, 1 unit of Aatll can be used to digest 8 µg of DNA in a 16 hour digest (+++).

    Extended activity was determined by performing the restriction endonuclease unit assay, using a 16 hour incubation in place of the standard 1 hour digestion. After the 16 hour digestion, extended activity enzymes (+++) required only 0.13 units to completely digest 1 µg of DNA. Intermediate activity enzymes required either 0.25 (++) or 0.50 (+) units for complete digestion over this extended incubation time. Finally, enzymes marked (-) required 1.0 unit for complete digestion, the same amount of enzyme required for a 1 hour digestion. Substrate DNA is defined in the unit assay for a particular product, which is provided on the individual product page.

    +++
    ++
    +
    Enzyme is active > 8 hours
    Enzyme is active 4-8 hours
    Enzyme is active 2-4 hours
    Not benefit from digesting over 1 hour

    EnzymeSuitability for Extended Digestions
    AatII+++
    AbaSI @25°C
    Acc65I+
    AccI+++
    AciI-
    AclI+
    AcuI-
    AfeI++
    AflII+++
    AflIII+++
    AgeI+
    AgeI-HF++
    AgeI-HF® RE-Mix®+
    AhdI+++
    AleI++
    AluI++
    AlwI+
    AlwNI+++
    ApaI @25°C+++
    ApaLI+++
    ApeKI @75°C+++
    ApoI @50°C+++
    AscI+++
    AscI RE-Mix®+++
    AseI+++
    AsiSI+++
    AvaI++
    AvaII++
    AvrII+++
    EnzymeBack to topSuitability for Extended Digestions
    BaeGI+
    BaeI @25°C+
    BamHI+
    BamHI-HF®+
    BanI+++
    BanII+
    BbsI++
    BbvCI+++
    BbvI-
    BccI+
    BceAI+++
    BcgI+
    BciVI-
    BclI @50°C+
    BcoDI+++
    BfaI+
    BfuAI @50°C++
    BfuCI-
    BglI+++
    BglII++
    BlpI+
    BmgBI-
    BmrI-
    BmtI+
    BmtI-HF®+++
    BpmI-
    Bpu10I+
    BpuEI-
    BsaAI++
    BsaBI @60°C+
    BsaHI+++
    BsaI-
    BsaI-HF®++
    BsaJI @60°C+++
    BsaWI @60°C+++
    BsaXI++
    BseRI+
    BseYI++
    BsgI+
    BsiEI @60°C++
    BsiHKAI @65°C-
    BsiWI @55°C+++
    BslI @55°C+++
    BsmAI @55°C++
    BsmBI @55°C+
    BsmFI @65°C+++
    BsmI @65°C+
    BsoBI+++
    Bsp1286I+
    BspCNI @25°C-
    BspDI++
    BspEI+++
    BspHI+++
    BspMI++
    BspQI @50°C-
    BsrBI+
    BsrDI @65°C+
    BsrFI+++
    BsrGI+++
    BsrI @65°C++
    BssHII @50°C+
    BssKI @60°C++
    BssSI++
    BstAPI @60°C++
    BstBI @65°C+++
    BstEII  @60°C-
    BstEII-HF®-
    BstEII-HF® RE-Mix®-
    BstNI @60°C-
    BstUI @60°C+++
    BstXI+++
    BstYI @60°C+++
    BstZ17I-
    Bsu36I+++
    BtgI+
    BtgZI @60°C-
    BtsCI @50°C+
    BtsI @55°C+
    BtsIMutI @55°C+
    EnzymeBack to topSuitability for Extended Digestions
    Cac8I++
    ClaI+
    CspCI+
    CviAII @25°C-
    CviKI-1-
    CviQI @25°C++
    EnzymeBack to topSuitability for Extended Digestions
    DdeI+++
    DpnI+++
    DpnII+++
    DraI++
    DraIII++
    DraIII-HF®+++
    DrdI+++
    DvuIII
    EnzymeBack to topSuitability for Extended Digestions
    EaeI+++
    EagI+++
    EagI-HF®-
    EarI+++
    EciI-
    Eco53kI++
    EcoNI+++
    EcoO109I+++
    EcoP15I-
    EcoRI+++
    EcoRI-HF®+++
    EcoRI-HF® RE-Mix®+++
    EcoRV+
    EcoRV-HF®+
    EcoRV-HF® RE-Mix®+
    EnzymeBack to topSuitability for Extended Digestions
    FatI @55°C+
    FauI @55°C-
    Fnu4HI++
    FokI-
    FseI-
    FspEI+++
    FspI+++
    EnzymeBack to topSuitability for Extended Digestions
    HaeII-
    HaeIII++
    HgaI-
    HhaI++
    HincII+++
    HindIII+++
    HindIII-HF®+++
    HinfI++
    HinP1I++
    HpaI++
    HpaII++
    HphI++
    Hpy166II+++
    Hpy188I-
    Hpy188III++
    Hpy99I-
    HpyAV-
    HpyCH4III+++
    HpyCH4IV++
    HpyCH4V++
    EnzymeBack to topSuitability for Extended Digestions
    I-CeuI++
    I-SceI++
    EnzymeBack to topSuitability for Extended Digestions
    KasI-
    KpnI++
    KpnI-HF®+
    KpnI-HF® RE-Mix®+
    EnzymeBack to topSuitability for Extended Digestions
    LpnPI-
    EnzymeBack to topSuitability for Extended Digestions
    MboI+
    MboII-
    MfeI++
    MfeI-HF®++
    MfeI-HF® RE-Mix®++
    MluCI-
    MluI+++
    MlyI-
    MmeI-
    MnlI++
    MscI+
    MseI+++
    MslI-
    MspA1I++
    MspI+
    MspJI+++
    MwoI @60°C+++
    EnzymeBack to topSuitability for Extended Digestions
    NaeI+
    NarI-
    Nb.BbvCI+++
    Nb.BsmI @65°C++
    Nb.BsrDI @65°C++
    Nb.BtsI++
    NciI+
    NcoI++
    NcoI-HF®++
    NcoI-HF® RE-Mix®++
    NdeI+++
    NgoMIV+++
    NheI++
    NheI-HF®+++
    NheI-HF® RE-Mix®++
    NlaIII+
    NlaIV+
    NmeAIII-
    NotI++
    NotI-HF®+++
    NotI-HF® RE-Mix® +++
    NruI+++
    NsiI++
    NspI++
    Nt.AlwI+++
    Nt.BbvCI+++
    Nt.BsmAI+++
    Nt.BspQI @50°C++
    Nt.BstNBI @55°C+
    Nt.CviPII-
    EnzymeBack to topSuitability for Extended Digestions
    PacI+++
    PacI RE-Mix®+++
    PaeR7I+++
    PciI++
    PflFI+++
    PflMI+
    PhoI @75°C+++
    PI-PspI @65°C+++
    PI-SceI+++
    PleI+
    PluTI
    PmeI-
    PmlI+
    PpuMI+++
    PshAI-
    PsiI++
    PspGI @75°C++
    PspOMI+++
    PspXI+++
    PstI+
    PstI-HF®+
    PvuI+++
    PvuI-HF®+++
    PvuII+++
    PvuII-HF®-
    EnzymeBack to topSuitability for Extended Digestions
    RsaI++
    RsrII++
    EnzymeBack to topSuitability for Extended Digestions
    SacI+++
    SacI-HF®+++
    SacII+++
    SalI+++
    SalI-HF®++
    SalI-HF® RE-Mix®+++
    SapI-
    Sau3AI+
    Sau96I++
    SbfI+
    SbfI-HF®-
    ScaI++
    ScaI-HF®++
    ScaI-HF® RE-Mix®++
    ScrFI++
    SexAI++
    SfaNI+
    SfcI-
    SfiI @50°C++
    SfoI-
    SgrAI-
    SmaI @25°C+++
    SmlI @55°C++
    SnaBI+
    SpeI++
    SpeI RE-Mix®++
    SpeI-HF®+
    SphI-HF®-
    SphI+++
    SspI++
    SspI-HF®+
    StuI++
    StyD4I+++
    StyI++
    StyI-HF®++
    SwaI @25°C++
    EnzymeBack to topSuitability for Extended Digestions
    TaqαI @65°C+
    TfiI @65°C++
    TliI @75°C++
    TseI @65°C+
    Tsp45I @65°C+
    Tsp509I @65°C+++
    TspMI @75°C+++
    TspRI @65°C+++
    Tth111I @65°C++
    EnzymeBack to topSuitability for Extended Digestions
    XbaI+++
    XbaI RE-Mix®+++
    XcmI+++
    XhoI+++
    XhoI RE-Mix®+++
    XmaI+
    XmnI++
    EnzymeBack to topSuitability for Extended Digestions
    ZraI+