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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
    AatII37+++
    AbaSI25
    Acc65I37+
    AccI37+++
    AciI37-
    AclI37+
    AcuI37-
    AfeI37++
    AflII37+++
    AflIII37+++
    AgeI37+
    AgeI-HF37++
    AgeI-HF® RE-Mix®37+
    AhdI37+++
    AleI37++
    AluI37++
    AlwI37+
    AlwNI37+++
    ApaI25+++
    ApaLI37+++
    ApeKI75+++
    ApoI50+++
    AscI37+++
    AscI RE-Mix®37+++
    AseI37+++
    AsiSI37+++
    AvaI37++
    AvaII37++
    AvrII37+++
    EnzymeBack to topSuitability for Extended Digestions
    BaeGI37+
    BaeI25+
    BamHI37+
    BamHI-HF®37+
    BanI37+++
    BanII37+
    BbsI37++
    BbvCI37+++
    BbvI37-
    BccI37+
    BceAI37+++
    BcgI37+
    BciVI37-
    BclI50+
    BcoDI37+++
    BfaI37+
    BfuAI50++
    BfuCI37-
    BglI37+++
    BglII37++
    BlpI37+
    BmgBI37-
    BmrI37-
    BmtI37+
    BmtI-HF®37+++
    BpmI37-
    Bpu10I37+
    BpuEI37-
    BsaAI37++
    BsaBI60+
    BsaHI37+++
    BsaI37-
    BsaI-HF®37++
    BsaJI60+++
    BsaWI60+++
    BsaXI37++
    BseRI37+
    BseYI37++
    BsgI37+
    BsiEI60++
    BsiHKAI65-
    BsiWI55+++
    BslI55+++
    BsmAI55++
    BsmBI55+
    BsmFI65+++
    BsmI65+
    BsoBI37+++
    Bsp1286I37+
    BspCNI25-
    BspDI37++
    BspEI37+++
    BspHI37+++
    BspMI37++
    BspQI50-
    BsrBI37+
    BsrDI65+
    BsrFI37+++
    BsrGI37+++
    BsrI65++
    BssHII50+
    BssKI60++
    BssSI37++
    BstAPI60++
    BstBI65+++
    BstEII 60-
    BstEII-HF®37-
    BstEII-HF® RE-Mix®37-
    BstNI60-
    BstUI60+++
    BstXI37+++
    BstYI60+++
    BstZ17I37-
    Bsu36I37+++
    BtgI37+
    BtgZI60-
    BtsCI50+
    BtsI55+
    BtsIMutI55+
    EnzymeBack to topSuitability for Extended Digestions
    Cac8I37++
    ClaI37+
    CspCI37+
    CviAII25-
    CviKI-137-
    CviQI25++
    EnzymeBack to topSuitability for Extended Digestions
    DdeI37+++
    DpnI37+++
    DpnII37+++
    DraI37++
    DraIII37++
    DraIII-HF®37+++
    DrdI37+++
    DvuIII
    EnzymeBack to topSuitability for Extended Digestions
    EaeI37+++
    EagI37+++
    EagI-HF®37-
    EarI37+++
    EciI37-
    Eco53kI37++
    EcoNI37+++
    EcoO109I37+++
    EcoP15I37-
    EcoRI37+++
    EcoRI-HF®37+++
    EcoRI-HF® RE-Mix®37+++
    EcoRV37+
    EcoRV-HF®37+
    EcoRV-HF® RE-Mix®37+
    EnzymeBack to topSuitability for Extended Digestions
    FatI55+
    FauI55-
    Fnu4HI37++
    FokI37-
    FseI37-
    FspEI37+++
    FspI37+++
    EnzymeBack to topSuitability for Extended Digestions
    HaeII37-
    HaeIII37++
    HgaI37-
    HhaI37++
    HincII37+++
    HindIII37+++
    HindIII-HF®37+++
    HinfI37++
    HinP1I37++
    HpaI37++
    HpaII37++
    HphI37++
    Hpy166II37+++
    Hpy188I37-
    Hpy188III37++
    Hpy99I37-
    HpyAV37-
    HpyCH4III37+++
    HpyCH4IV37++
    HpyCH4V37++
    EnzymeBack to topSuitability for Extended Digestions
    I-CeuI37++
    I-SceI37++
    EnzymeBack to topSuitability for Extended Digestions
    KasI37-
    KpnI37++
    KpnI-HF®37+
    KpnI-HF® RE-Mix®37+
    EnzymeBack to topSuitability for Extended Digestions
    LpnPI37-
    EnzymeBack to topSuitability for Extended Digestions
    MboI37+
    MboII37-
    MfeI37++
    MfeI-HF®37++
    MfeI-HF® RE-Mix®37++
    MluCI37-
    MluI37+++
    MlyI37-
    MmeI37-
    MnlI37++
    MscI37+
    MseI37+++
    MslI37-
    MspA1I37++
    MspI37+
    MspJI37+++
    MwoI60+++
    EnzymeBack to topSuitability for Extended Digestions
    NaeI37+
    NarI37-
    Nb.BbvCI37+++
    Nb.BsmI65++
    Nb.BsrDI65++
    Nb.BtsI37++
    NciI37+
    NcoI37++
    NcoI-HF®37++
    NcoI-HF® RE-Mix®37++
    NdeI37+++
    NgoMIV37+++
    NheI37++
    NheI-HF®37+++
    NheI-HF® RE-Mix®37++
    NlaIII37+
    NlaIV37+
    NmeAIII37-
    NotI37++
    NotI-HF®37+++
    NotI-HF® RE-Mix® 37+++
    NruI37+++
    NsiI37++
    NspI37++
    Nt.AlwI37+++
    Nt.BbvCI37+++
    Nt.BsmAI37+++
    Nt.BspQI50++
    Nt.BstNBI55+
    Nt.CviPII37-
    EnzymeBack to topSuitability for Extended Digestions
    PacI37+++
    PacI RE-Mix®37+++
    PaeR7I37+++
    PciI37++
    PflFI37+++
    PflMI37+
    PhoI75+++
    PI-PspI65+++
    PI-SceI37+++
    PleI37+
    PluTI37
    PmeI37-
    PmlI37+
    PpuMI37+++
    PshAI37-
    PsiI37++
    PspGI75++
    PspOMI37+++
    PspXI37+++
    PstI37+
    PstI-HF®37+
    PvuI37+++
    PvuI-HF®37+++
    PvuII37+++
    PvuII-HF®37-
    EnzymeBack to topSuitability for Extended Digestions
    RsaI37++
    RsrII37++
    EnzymeBack to topSuitability for Extended Digestions
    SacI37+++
    SacI-HF®37+++
    SacII37+++
    SalI37+++
    SalI-HF®37++
    SalI-HF® RE-Mix®37+++
    SapI37-
    Sau3AI37+
    Sau96I37++
    SbfI37+
    SbfI-HF®37-
    ScaI37++
    ScaI-HF®37++
    ScaI-HF® RE-Mix®37++
    ScrFI37++
    SexAI37++
    SfaNI37+
    SfcI37-
    SfiI50++
    SfoI37-
    SgrAI37-
    SmaI25+++
    SmlI55++
    SnaBI37+
    SpeI37++
    SpeI RE-Mix®37++
    SpeI-HF®37+
    SphI-HF®37-
    SphI37+++
    SspI37++
    SspI-HF®37+
    StuI37++
    StyD4I37+++
    StyI37++
    StyI-HF®37++
    SwaI25++
    EnzymeBack to topSuitability for Extended Digestions
    TaqαI65+
    TfiI65++
    TliI75++
    TseI65+
    Tsp45I65+
    Tsp509I65+++
    TspMI75+++
    TspRI65+++
    Tth111I65++
    EnzymeBack to topSuitability for Extended Digestions
    XbaI37+++
    XbaI RE-Mix®37+++
    XcmI37+++
    XhoI37+++
    XhoI RE-Mix®37+++
    XmaI37+
    XmnI37++
    EnzymeBack to topSuitability for Extended Digestions
    ZraI37+