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 §11
AhdI11
AlwNI21
ApaI11
ApoI §11
AseI0.31
AvaI101
AvrII11
BaeI31
BamHI §31
BanII11
BglII81
BpmI11
BsaAI201
BsaI §21
BsaWI31
BsaXI21
BsgI11
BsmI11
BspDI11
BspEI11
BspMI**1
BspQI31
BsrFI21
BsrGI §11
BssHII41
BstZ17I-HF®11
BtgI51
ClaI51
EagI §101
EcoNI31
EcoO109I81
EcoRI §31
EcoRV §11
HincII41
HindIII §51
KasI41
KpnI §21
MluI §21
NarI101
NcoI §11
NdeI31
NheI §51
NmeAIII**1
NruI §11
NsiI §11
PciI31
PsiI31
PstI §11
PvuI §21
PvuII §21
SacI §51
SalI §101
SapI11
ScaI §201
SmaI11
SnaBI11
SpeI §11
SphI §31
SspI §41
StuI31
StyI §41
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.
§ A HF version of this enzyme is available.