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Restriction Endonucleases

The Leader in the Discovery and Production of Restriction Enzymes

With over 45 years of offering restriction enzymes to the research community, NEB has earned the reputation of being a leader in enzyme technologies. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled performance.

NEB scientists continue to improve its portfolio of restriction enzymes, as well as explore their utility in new technologies. As a result, NEB scientists continue to publish scientific papers and to be awarded grants in this area. With the industry’s largest research and development group dedicated to restriction enzymes, we are proud to have been there first: the first to commercialize a recombinant enzyme, the first to introduce a nicking enzyme. In addition, NEB has an ongoing history of innovation by engineering restriction enzymes with altered specificities and improved performance. Through continued research in these areas, we are committed to driving the innovations that allow us to offer maximum convenience and performance. 

For details on NEB’s quality controls for restriction endonucleases, visit our Restriction Enzyme Quality page.

[DNA thumbnail] Cut Smarter with Restriction Enzymes from NEB®


All of NEB's Restriction enzymes have transitioned to a new buffer system. Visit NEBCutSmart.com for further details.


Convenience

  • >210 restriction enzymes are 100% active in a single buffer – rCutSmart™ Buffer.
  • >180 restriction enzymes are Time-Saver qualified, meaning you can digest DNA in 5-15 minutes, or digest DNA safely overnight.
  • Choose from >265 restriction enzymes, the largest selection commercially available.

[DNA thumbnail] View the Activity/Performance Chart for NEB® Restriction Enzymes


Performance

  • Choose a High-Fidelity (HF®) restriction enzyme, which has been engineered for reduced star activity, rapid digestion (5-15 minutes) and 100% activity in rCutSmart Buffer. A vial of 6X Purple Loading Dye is included with most restriction enzymes.
  • All of our restriction enzymes undergo stringent quality control testing, ensuring the highest levels of purity and lot-to-lot consistency.


    Physical Purity Enzymes are evaluated for purity to ensure the highest levels of purity and the absence of contaminating proteins.
    DNA Contamination Enzymes are screened by qPCR to ensure no contaminating genomic DNA is present. The specification for this assay is less than one E.coli genome per sample.
    Exonuclease Activity Using radioactively labelled DNA substrate and/or state-of-the-art capillary electrophoresis-based assays with fluorescently-labelled substrates, NEB is able to detect very low levels of exonuclease activity.
    Endonuclease Activity To ensure that there are no contaminating enzymes that could cause nicking or non-specific nuclease degradation, reagents are incubated with supercoiled plasmid DNA for 4 hours to demonstrate the absence of endonuclease contamination. 
    Non-Specific DNase Activity Enzymes are incubated overnight with Lambda DNA to confirm that there is no additional non-specific nuclease activity present. 
    Cloning QC (Ligation and Re-cutting) A DNA template is over-digested by the appropriate restriction enzyme and the percentage of DNA fragments ligated and re-cut are determined by agarose gel electrophoresis.
    Cloning QC (Blue-white Screening) A DNA plasmid is over-digested by the appropriate restriction enzyme and the linearized plasmid DNA is ligated and transformed into an E.coli strain with greater than 99% correct transformants, as determined by a blue-white screen. 

Use Enzyme Finder to select restriction enzyme by name, sequence, overhang or type. 

Restriction enzyme infographic

Cut Smarter with Restriction Enzymes from NEB

With over 45 years of offering restriction enzymes to the research community, NEB has earned the reputation of being a leader in enzyme technologies. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled performance.

Convenience

  • A vial of 6X Purple Load Dye is included with most restriction enzymes.
  • Over 210 restriction enzymes are 100% active in a single buffer – rCutSmart™ Buffer. 
  • >180 restriction enzymes are Time-Saver qualified, meaning you can digest DNA in 5-15 minutes, or digest DNA safely overnight.
  • Choose from >265 restriction enzymes, the largest selection commercially available.



Performance

  • Choose a High-Fidelity (HF®) restriction enzyme, which has been engineered for reduced star activity, rapid digestion (5-15 minutes) and 100% activity in rCutSmart Buffer. A vial of 6X Purple Load Dye is included with every HF restriction enzyme.
  • All of our restriction enzymes undergo stringent quality control testing, ensuring the highest levels of purity and lot-to-lot consistency.


Use Enzyme Finder to select restriction enzymes by name, sequence, overhang or type.


Choose Product:

Restriction Endonucleases

WarmStart® Nt.BstNBI

Restriction Endonucleases: A

AatII

Acc65I

AccI

AciI

AclI

AcuI

AfeI

AflII

AflIII

AgeI-HF®

AhdI

AleI-v2

AluI

AlwI

AlwNI

ApaI

ApaLI

ApeKI

ApoI-HF®

AscI

AseI

AsiSI

AvaI

AvaII

AvrII

Restriction Endonucleases B

BaeGI

BaeI

BamHI

BamHI-HF®

BanI

BanII

BbsI

BbsI-HF®

BbvCI

BbvI

BccI

BceAI

BcgI

BciVI

BclI

BclI-HF®

BcoDI

BfaI

BfuAI

BglI

BglII

BlpI

BmgBI

BmrI

BmtI-HF®

BpmI

Bpu10I

BpuEI

BsaAI

BsaBI

BsaHI

BsaI-HF®v2

BsaJI

BsaWI

BsaXI

BseRI

BseYI

BsgI

BsiEI

BsiHKAI

BsiWI

BsiWI-HF®

BslI

BsmAI

BsmBI-v2

BsmFI

BsmI

BsoBI

Bsp1286I

BspCNI

BspDI

BspEI

BspHI

BspMI

BspQI

BspQI-HF®

BsrBI

BsrDI

BsrFI-v2

BsrGI-HF®

BsrI

BssHII

BssSI-v2

BstAPI

BstBI

BstEII-HF®

BstNI

BstUI

BstXI

BstYI

BstZ17I-HF®

Bsu36I

BtgI

BtgZI

BtsCI

BtsIMutI

BtsI-v2

Restriction Endonucleases C G

Cac8I

ClaI

CspCI

CviKI-1

CviQI

DdeI

DpnI

DpnII

DraI

DraIII-HF®

DrdI

EaeI

EagI-HF®

EarI

EciI

Eco53kI

EcoNI

EcoO109I

EcoP15I

EcoRI

EcoRI-HF®

EcoRV

EcoRV-HF®

Esp3I

FatI

FauI

Fnu4HI

FokI

FseI

FspI

Restriction Endonucleases H M

HaeII

HaeIII

HgaI

HhaI

HincII

HindIII

HindIII-HF®

HinfI

HinP1I

HpaI

HpaII

HphI

Hpy166II

Hpy188I

Hpy188III

Hpy99I

HpyAV

HpyCH4III

HpyCH4IV

HpyCH4V

I-CeuI

I-SceI

KasI

KpnI-HF®

MboI

MboII

MfeI-HF®

MluCI

MluI-HF®

MlyI

MmeI

MnlI

MscI

MseI

MslI

MspA1I

MspI

MspJI

MwoI

Restriction Endonucleases: N-O

NaeI

NarI

Nb.BbvCI

Nb.BsmI

Nb.BsrDI

Nb.BssSI

Nb.BtsI

NciI

NcoI

NcoI-HF®

NdeI

NgoMIV

NheI-HF®

NlaIII

NlaIV

NmeAIII

NotI

NotI-HF®

NruI-HF®

NsiI

NsiI-HF®

NspI

Nt.AlwI

Nt.BbvCI

Nt.BsmAI

Nt.BspQI

Nt.BstNBI

Nt.CviPII

WarmStart® Nt.BstNBI

Restriction Endonucleases P R

PacI

PaeR7I

PaqCI®

PciI

PflFI

PflMI

PI-PspI

PI-SceI

PleI

PluTI

PmeI

PmlI

PpuMI

PshAI

PsiI-v2

PspGI

PspOMI

PspXI

PstI

PstI-HF®

PvuI-HF®

PvuII

PvuII-HF®

RsaI

RsrII

Restriction Endonucleases S

SacI-HF®

SacII

SalI

SalI-HF®

SapI

Sau3AI

Sau96I

SbfI-HF®

ScaI-HF®

ScrFI

SexAI

SfaNI

SfcI

SfiI

SfoI

SgrAI

SmaI

SmlI

SnaBI

SpeI-HF®

SphI

SphI-HF®

SrfI

SspI-HF®

StuI

StyD4I

StyI-HF®

SwaI

Restriction Endonucleases T Z

TaqI-v2

TfiI

TseI

Tsp45I

TspMI

TspRI

Tth111I

XbaI

XcmI

XhoI

XmaI

XmnI

ZraI

HF, Nicking, Master Mix, Time-Saver & Other

AatII

Acc65I

AccI

AciI

AclI

AcuI

AflII

AgeI-HF®

AhdI

AluI

AlwNI

ApaI

ApaLI

ApeKI

ApoI-HF®

AscI

AseI

AvaI

AvaII

AvrII

BaeGI

BaeI

BamHI

BamHI-HF®

BbsI

BbsI-HF®

BbvI

BciVI

BclI

BclI-HF®

BcoDI

BfuAI

BglI

BglII

BlpI

BmgBI

BmtI-HF®

BpmI

BpuEI

BsaAI

BsaHI

BsaI-HF®v2

BsaWI

BsaXI

BseRI

BsgI

BsiEI

BsiWI

BsiWI-HF®

BslI

BsmAI

BsmBI-v2

BsmI

BsoBI

Bsp1286I

BspCNI

BspEI

BspHI

BspQI

BspQI-HF®

BsrBI

BsrDI

BsrGI-HF®

BsrI

BssHII

BstBI

BstEII-HF®

BstNI

BstUI

BstXI

BstYI

BstZ17I-HF®

Bsu36I

BtgI

BtsCI

Cac8I

ClaI

CspCI

CviQI

DdeI

DpnI

DpnII

DraI

DraIII-HF®

DrdI

EagI-HF®

EarI

Eco53kI

EcoNI

EcoO109I

EcoP15I

EcoRI

EcoRI-HF®

EcoRV

EcoRV-HF®

Esp3I

Fnu4HI

FseI

FspI

HaeIII

HhaI

HincII

HindIII-HF®

HinfI

HinP1I

HpaII

HphI

Hpy166II

HpyAV

HpyCH4IV

HpyCH4V

I-CeuI

I-SceI

KpnI-HF®

MboI

MfeI-HF®

MluCI

MluI-HF®

MlyI

MmeI

MnlI

MseI

MslI

MspA1I

MspI

MspJI

MwoI

Nb.BbvCI

Nb.BsmI

Nb.BsrDI

Nb.BssSI

Nb.BtsI

NciI

NcoI

NcoI-HF®

NdeI

NgoMIV

NheI-HF®

NlaIII

NotI

NotI-HF®

NruI-HF®

NsiI

NsiI-HF®

NspI

Nt.AlwI

Nt.BbvCI

Nt.BsmAI

Nt.BspQI

Nt.BstNBI

Nt.CviPII

PacI

PaeR7I

PflFI

PflMI

PI-PspI

PI-SceI

PmeI

PmlI

PpuMI

PshAI

PstI

PstI-HF®

PvuI-HF®

PvuII

PvuII-HF®

RsaI

SacI-HF®

SacII

SalI

SalI-HF®

SapI

SbfI-HF®

ScaI-HF®

SfiI

SfoI

SmaI

SpeI-HF®

SphI-HF®

SspI-HF®

StuI

StyD4I

StyI-HF®

SwaI

TaqI-v2

TfiI

TseI

TspMI

TspRI

Tth111I

WarmStart® Nt.BstNBI

XbaI

XhoI

XmaI

XmnI

Restriction Endonuclease Buffers & Diluents

Diluent A (with rAlbumin)

Diluent B (with rAlbumin)

Diluent C (with rAlbumin)

NEBuffer™ 1

NEBuffer™ 2

NEBuffer™ 3

NEBuffer™ 4

NEBuffer™ r2.1

NEBuffer™ r3.1

NEBuffer™ Set (EcoRI/SspI, DpnII)

NEBuffer™ Set (r1.1, r2.1, r3.1 and rCutSmart™)

rCutSmart™ Buffer

Recombinant Albumin, Molecular Biology Grade

Recombinant Albumin, Molecular Biology Grade (Glycerol-free)

S-adenosylmethionine (SAM)

Restriction Endonucleases: A

AatII

Acc65I

AccI

AciI

AclI

AcuI

AfeI

AflII

AflIII

AgeI-HF®

AhdI

AleI-v2

AluI

AlwI

AlwNI

ApaI

ApaLI

ApeKI

ApoI-HF®

AscI

AseI

AsiSI

AvaI

AvaII

AvrII

Restriction Endonucleases B

BaeGI

BaeI

BamHI

BamHI-HF®

BanI

BanII

BbsI

BbsI-HF®

BbvCI

BbvI

BccI

BceAI

BcgI

BciVI

BclI

BclI-HF®

BcoDI

BfaI

BfuAI

BglI

BglII

BlpI

BmgBI

BmrI

BmtI-HF®

BpmI

Bpu10I

BpuEI

BsaAI

BsaBI

BsaHI

BsaI-HF®v2

BsaJI

BsaWI

BsaXI

BseRI

BseYI

BsgI

BsiEI

BsiHKAI

BsiWI

BsiWI-HF®

BslI

BsmAI

BsmBI-v2

BsmFI

BsmI

BsoBI

Bsp1286I

BspCNI

BspDI

BspEI

BspHI

BspMI

BspQI

BspQI-HF®

BsrBI

BsrDI

BsrFI-v2

BsrGI-HF®

BsrI

BssHII

BssSI-v2

BstAPI

BstBI

BstEII-HF®

BstNI

BstUI

BstXI

BstYI

BstZ17I-HF®

Bsu36I

BtgI

BtgZI

BtsCI

BtsIMutI

BtsI-v2

Restriction Endonucleases C G

Cac8I

ClaI

CspCI

CviKI-1

CviQI

DdeI

DpnI

DpnII

DraI

DraIII-HF®

DrdI

EaeI

EagI-HF®

EarI

EciI

Eco53kI

EcoNI

EcoO109I

EcoP15I

EcoRI

EcoRI-HF®

EcoRV

EcoRV-HF®

Esp3I

FatI

FauI

Fnu4HI

FokI

FseI

FspI

Restriction Endonucleases H M

HaeII

HaeIII

HgaI

HhaI

HincII

HindIII

HindIII-HF®

HinfI

HinP1I

HpaI

HpaII

HphI

Hpy166II

Hpy188I

Hpy188III

Hpy99I

HpyAV

HpyCH4III

HpyCH4IV

HpyCH4V

I-CeuI

I-SceI

KasI

KpnI-HF®

MboI

MboII

MfeI-HF®

MluCI

MluI-HF®

MlyI

MmeI

MnlI

MscI

MseI

MslI

MspA1I

MspI

MspJI

MwoI

Restriction Endonucleases: N-O

NaeI

NarI

Nb.BbvCI

Nb.BsmI

Nb.BsrDI

Nb.BssSI

Nb.BtsI

NciI

NcoI

NcoI-HF®

NdeI

NgoMIV

NheI-HF®

NlaIII

NlaIV

NmeAIII

NotI

NotI-HF®

NruI-HF®

NsiI

NsiI-HF®

NspI

Nt.AlwI

Nt.BbvCI

Nt.BsmAI

Nt.BspQI

Nt.BstNBI

Nt.CviPII

WarmStart® Nt.BstNBI

Restriction Endonucleases P R

PacI

PaeR7I

PaqCI®

PciI

PflFI

PflMI

PI-PspI

PI-SceI

PleI

PluTI

PmeI

PmlI

PpuMI

PshAI

PsiI-v2

PspGI

PspOMI

PspXI

PstI

PstI-HF®

PvuI-HF®

PvuII

PvuII-HF®

RsaI

RsrII

Restriction Endonucleases S

SacI-HF®

SacII

SalI

SalI-HF®

SapI

Sau3AI

Sau96I

SbfI-HF®

ScaI-HF®

ScrFI

SexAI

SfaNI

SfcI

SfiI

SfoI

SgrAI

SmaI

SmlI

SnaBI

SpeI-HF®

SphI

SphI-HF®

SrfI

SspI-HF®

StuI

StyD4I

StyI-HF®

SwaI

Restriction Endonucleases T Z

TaqI-v2

TfiI

TseI

Tsp45I

TspMI

TspRI

Tth111I

XbaI

XcmI

XhoI

XmaI

XmnI

ZraI

HF, Nicking, Master Mix, Time-Saver & Other

High-Fidelity (HF®) Restriction Endonucleases

AgeI-HF®

ApoI-HF®

BamHI-HF®

BbsI-HF®

BclI-HF®

BmtI-HF®

BsaI-HF®v2

BsiWI-HF®

BspQI-HF®

BsrGI-HF®

BstEII-HF®

BstZ17I-HF®

DraIII-HF®

EagI-HF®

EcoRI-HF®

EcoRV-HF®

HindIII-HF®

KpnI-HF®

MfeI-HF®

MluI-HF®

NcoI-HF®

NheI-HF®

NotI-HF®

NruI-HF®

NsiI-HF®

PstI-HF®

PvuI-HF®

PvuII-HF®

SacI-HF®

SalI-HF®

SbfI-HF®

ScaI-HF®

SpeI-HF®

SphI-HF®

SspI-HF®

StyI-HF®

Homing Endonucleases

I-CeuI

I-SceI

PI-PspI

PI-SceI

Nicking Endonucleases

Nb.BbvCI

Nb.BsmI

Nb.BsrDI

Nb.BssSI

Nb.BtsI

Nt.AlwI

Nt.BbvCI

Nt.BsmAI

Nt.BspQI

Nt.BstNBI

Nt.CviPII

WarmStart® Nt.BstNBI

Restriction Enzymes for Epigenetic Analysis

DpnI

DpnII

HpaII

MspI

MspJI

Time-Saver™ Qualified Restriction Enzymes

AatII

Acc65I

AccI

AciI

AclI

AcuI

AflII

AgeI-HF®

AhdI

AluI

AlwNI

ApaI

ApaLI

ApeKI

ApoI-HF®

AscI

AseI

AvaI

AvaII

AvrII

BaeGI

BaeI

BamHI

BamHI-HF®

BbsI

BbsI-HF®

BbvI

BciVI

BclI

BclI-HF®

BcoDI

BfuAI

BglI

BglII

BlpI

BmgBI

BmtI-HF®

BpmI

BpuEI

BsaAI

BsaHI

BsaI-HF®v2

BsaWI

BsaXI

BseRI

BsgI

BsiEI

BsiWI

BsiWI-HF®

BslI

BsmAI

BsmBI-v2

BsmI

BsoBI

Bsp1286I

BspCNI

BspEI

BspHI

BspQI

BspQI-HF®

BsrBI

BsrDI

BsrI

BssHII

BstBI

BstEII-HF®

BstNI

BstUI

BstXI

BstYI

BstZ17I-HF®

Bsu36I

BtgI

BtsCI

Cac8I

ClaI

CspCI

CviQI

DdeI

DpnI

DpnII

DraI

DraIII-HF®

DrdI

EagI-HF®

EarI

Eco53kI

EcoNI

EcoO109I

EcoP15I

EcoRI

EcoRI-HF®

EcoRV

EcoRV-HF®

Esp3I

Fnu4HI

FseI

FspI

HaeIII

HhaI

HincII

HindIII-HF®

HinfI

HinP1I

HpaII

HphI

Hpy166II

HpyAV

HpyCH4IV

HpyCH4V

KpnI-HF®

MboI

MfeI-HF®

MluCI

MlyI

MmeI

MnlI

MseI

MslI

MspA1I

MspI

MwoI

NciI

NcoI

NcoI-HF®

NdeI

NgoMIV

NheI-HF®

NlaIII

NotI

NotI-HF®

NsiI

NspI

PacI

PaeR7I

PflFI

PflMI

PmeI

PmlI

PpuMI

PshAI

PstI

PstI-HF®

PvuI-HF®

PvuII

PvuII-HF®

RsaI

SacI-HF®

SacII

SalI

SalI-HF®

SapI

SbfI-HF®

ScaI-HF®

SfiI

SfoI

SmaI

SpeI-HF®

SphI-HF®

SspI-HF®

StuI

StyD4I

StyI-HF®

SwaI

TaqI-v2

TfiI

TseI

TspMI

TspRI

Tth111I

XbaI

XhoI

XmaI

XmnI

Restriction Endonuclease Buffers & Diluents

Diluent A (with rAlbumin)

Diluent B (with rAlbumin)

Diluent C (with rAlbumin)

NEBuffer™ 1

NEBuffer™ 2

NEBuffer™ 3

NEBuffer™ 4

NEBuffer™ r2.1

NEBuffer™ r3.1

NEBuffer™ Set (EcoRI/SspI, DpnII)

NEBuffer™ Set (r1.1, r2.1, r3.1 and rCutSmart™)

rCutSmart™ Buffer

Recombinant Albumin, Molecular Biology Grade

Recombinant Albumin, Molecular Biology Grade (Glycerol-free)

S-adenosylmethionine (SAM)


Choose Type:

Restriction Endonucleases includes these subcategories:
Restriction Endonucleases: A
Restriction Endonucleases: B
Restriction Endonucleases: C-G
Restriction Endonucleases: H-M
Restriction Endonucleases: N-O
Restriction Endonucleases: P-R
Restriction Endonucleases: S
Restriction Endonucleases: T-Z
HF®, Nicking, Master Mix, Time-Saver & Other
Restriction Endonuclease Buffers & Diluents
FAQs for Restriction Endonucleases
Protocols for Restriction Endonucleases
Application Notes for Restriction Endonucleases
    Publications related to Restriction Endonucleases
    • Shah, S., Sanchez, J., Stewart, A., et al. (2015) Probing the Run-On Oligomer of Activated SgrAI Bound to DNA PLoS One; 10(4), PubMedID: 25880668, DOI: 10.1371/journal.pone.0124783.
    • Roberts, R.J., Vincze, T., Posfai, J., Macelis, D. (2015) REBASE - A database for DNA restriction and modification: enzymes, genes and genomes Nucleic Acids Res; 43, D298-D299. PubMedID: 25378308
    • Fomenkov, A., Lunnen, K.D., Zhu, Z., Anton, B.P., Wilson, G.G., Vincze, T. and Roberts, R.J. (2015) Complete genome sequence and methylome analysis of bacillus strain x1 Genome Announc; 3(1), PubMedID: 25700417
    • Loenen, W.A., Raleigh, E.A. (2014) The other face of restriction: modification-dependent enzymes. Nucleic Acids Res; 42, 56-69. PubMedID: 23990325, DOI: doi: 10.1093/nar/gkt847
    • Roberts, R.J., Vincze, T., Posfai, J., Macelis, D. (2014) REBASE - A database for DNA restriction and modification: enzymes, genes and genomes Nucleic Acids Res; PubMedID: 25378308
    • Kamps-Hughes, N., Quimby, A., Zhu, Z., Johnson, E.A. (2013) Massively parallel characterization of restriction endonucleases Nucleic Acids Res ; 41(11), e119. PubMedID: 23605040, DOI: 10.1093/nar/gkt257
    • Fu YB, Peterson G. W., Dong Y (2016) Increasing Genome Sampling and Improving SNP Genotyping for Genotyping-by-Sequencing with New Combinations of Restriction Enzymes G3 (Bethesda); 6:4, 845-846. PubMedID: 26818077
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