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