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Conventional Enzymes

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AanI

PsiI
AanI (PsiI)
Optimal Buffer: Tango
TTA↓TAA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R a
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 65°C

AarI

AarI
Optimal Buffer: Unique (+oligo)
CACCTGC(4/8)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X (+oligo)
G unique
O twox
R a
1X Tango NR
2X Tango NR
Optimal Temperature 37°C
Double Digest 0-20 (+oligo)
Overnight 1
Inactivation 65°C

AatII

AatII
Optimal Buffer: Tango
GACGT↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R a
1X Tango 50-100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.3
Inactivation 65°C

Acc65I

Asp718I
Acc65I (Asp718I)
Optimal Buffer: O
G↓GTACC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R a
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.3
Inactivation 65°C

AdeI

DraIII
AdeI (DraIII)
Optimal Buffer: G
CACNNN↓GTG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X* or 2X
G green
O onex twox
R a
1X Tango 0-20
2X Tango 100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.5
Inactivation NO

AjiI

BmgBI
AjiI (BmgBI)
Optimal Buffer: Unique
CAC↓GTC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X*
G unique
O twox
R a
1X Tango NR
2X Tango NR
Optimal Temperature 37°C
Double Digest 20-50*
Overnight 0.5
Inactivation 65°C

AjuI

AjuI
Optimal Buffer: R (+SAM)
↓(7/12)GAA(N)7TTGG(11/6)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X (+SAM)
G red
O onex twox
R a
1X Tango 0-20 (+SAM)
2X Tango 50-100 (+SAM)
Optimal Temperature 37°C
Double Digest 20-50 (+SAM)
Overnight 0.5
Inactivation 65°C

AlfI

AlfI
Optimal Buffer: R (+SAM)
↓(10/12)GCA(N)6TGC(12/10)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X (+SAM)
G red
O twox
R a
1X Tango 0-20 (+SAM)
2X Tango 0-20 (+SAM)
Optimal Temperature 37°C
Double Digest 0-20 (+SAM)
Overnight 1
Inactivation 65°C

AloI

AloI
Optimal Buffer: R
↓(7/12-13)GAAC(N)6TCC(12-13/7)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R a
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 30°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

AluI

AluI
Optimal Buffer: Tango
AG↓CT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R a
1X Tango 50-100
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

Alw21I

BsiHKAI
Alw21I (BsiHKAI)
Optimal Buffer: O
GWGCW↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R a
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 65°C

Alw26I

BsmAI
Alw26I (BsmAI)
Optimal Buffer: Tango
GTCTC(1/5)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R a
1X Tango 50-100
2X Tango 100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

Alw44I

ApaLI
Alw44I (ApaLI)
Optimal Buffer: Tango
G↓TGCAC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R a
1X Tango 50-100
2X Tango 100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

ApaI

ApaI
Optimal Buffer: B
GGGCC↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R a
1X Tango 100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

BamHI

BamHI
Optimal Buffer: Unique
G↓GATCC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X* or 2X
G unique
O onex twox
R b
1X Tango 20-50*
2X Tango 100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.5
Inactivation 80°C (10 u)

BauI

BssSI
BauI (BssSI)
Optimal Buffer: Tango
CACGAG(-5/-1)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

BclI

BclI
Optimal Buffer: G
T↓GATCA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X* or 2X
G green
O onex twox
R b
1X Tango 20-50
2X Tango 100
Optimal Temperature 55°C
Double Digest 20-50
Overnight 0.1
Inactivation 80°C (10 u)

BcnI

NciI
BcnI (NciI)
Optimal Buffer: Tango
CC↓SGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.2
Inactivation 65°C

BcuI

SpeI
BcuI (SpeI)
Optimal Buffer: Tango
A↓CTAGT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R b
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation NO

BfmI

SfcI
BfmI (SfcI)
Optimal Buffer: Tango
C↓TRYAG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 1
Inactivation 65°C

BfuI

BciVI
BfuI (BciVI)
Optimal Buffer: Unique
GTATCC(6/5)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X*
G unique
O twox
R b
1X Tango NR
2X Tango NR
Optimal Temperature 37°C
Double Digest 0-20
Overnight 1
Inactivation 80°C

BglI

BglI
Optimal Buffer: O
GCCNNNN↓NGGC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R b
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 65°C

BglII

BglII
Optimal Buffer: O
A↓GATCT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R b
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation NO

Bme1390I

ScrFI
Bme1390I (ScrFI)
Optimal Buffer: O
CC↓NGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R b
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 100
Overnight 0.2
Inactivation 80°C

BoxI

PshAI
BoxI (PshAI)
Optimal Buffer: Tango
GACNN↓NNGTC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 80°C

BpiI

BbsI
BpiI (BbsI)
Optimal Buffer: G
GAAGAC(2/6)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G green
O onex twox
R b
1X Tango 20-50
2X Tango 100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.3
Inactivation 65°C

BplI

BplI
Optimal Buffer: Tango(+SAM)
↓(8/13)GAG(N)5CTC(13/8)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X (+SAM)
G yellow
O onex
R b
1X Tango 0-20 (+SAM)
2X Tango 20-50 (+SAM)
Optimal Temperature 37°C
Double Digest 0-20 (+SAM)
Overnight 0.3
Inactivation 65°C

Bpu10I

Bpu10I
Optimal Buffer: Unique
CCTNAGC(-5/-2)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X* or 2X*
G unique
O onex twox
R b
1X Tango 0-20
2X Tango 20-50*
Optimal Temperature 37°C
Double Digest 50-100*
Overnight 0.2
Inactivation 80°C

Bpu1102I

BlpI
Bpu1102I (BlpI)
Optimal Buffer: Tango
GC↓TNAGC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation 80°C

BseDI

BsaJI
BseDI (BsaJI)
Optimal Buffer: Tango
C↓CNNGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 50-100
2X Tango 20-50
Optimal Temperature 55°C
Double Digest 0-20
Overnight 0.2
Inactivation 80°C

BseGI

BtsCI
BseGI (BtsCI)
Optimal Buffer: Tango
GGATG(2/0)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 55°C
Double Digest 20-50
Overnight 0.2
Inactivation 80°C

BseJI

BsaBI
BseJI (BsaBI)
Optimal Buffer: O
GATNN↓NNATC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X*
G orange
O twox
R b
1X Tango NR
2X Tango 100*
Optimal Temperature 65°C
Double Digest 100
Overnight 0.1
Inactivation NO

BseLI

BslI
BseLI (BslI)
Optimal Buffer: Tango
CCNNNNN↓NNGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 20-50
2X Tango 100
Optimal Temperature 55°C
Double Digest 50-100
Overnight 0.1
Inactivation NO

BseMI

BsrDI
BseMI (BsrDI)
Optimal Buffer: R
GCAATG(2/0)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R b
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 55°C
Double Digest 0-20
Overnight 0.3
Inactivation 80°C

BseMII

BspCNI
BseMII (BspCNI)
Optimal Buffer: Tango(+SAM)
CTCAG(10/8)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X (+SAM)
G yellow
O onex twox
R b
1X Tango 50-100 (+SAM)
2X Tango 50-100 (+SAM)
Optimal Temperature 55°C
Double Digest 50-100 (+SAM)
Overnight 0.5
Inactivation 80°C

BseNI

BsrI
BseNI (BsrI)
Optimal Buffer: B
ACTGG(1/-1)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G blue
O onex twox
R b
1X Tango 100
2X Tango 20-50
Optimal Temperature 65°C
Double Digest 0-20
Overnight 0.1
Inactivation 80°C

BseSI

Bme1580I
BseSI (Bme1580I)
Optimal Buffer: G
GKGCM↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G green
O onex
R b
1X Tango 20-50
2X Tango 100
Optimal Temperature 55°C
Double Digest 0-20
Overnight 0.1
Inactivation 80°C (10 u)

BseXI

BbvI
BseXI (BbvI)
Optimal Buffer: Unique
GCAGC(8/12)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B NR
G unique
O 0
R b
1X Tango NR
2X Tango NR
Optimal Temperature 65°C
Double Digest NR
Overnight 0.3
Inactivation 80°C

Bsh1236I

BstUI
Bsh1236I (BstUI)
Optimal Buffer: R
CG↓CG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R b
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 65°C

Bsh1285I

BsiEI
Bsh1285I (BsiEI)
Optimal Buffer: G
CGRY↓CG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G green
O twox
R b
1X Tango 20-50
2X Tango 100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.2
Inactivation 80°C

BshNI

BanI
BshNI (BanI)
Optimal Buffer: O
G↓GYRCC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R b
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.3
Inactivation 65°C

BshTI

AgeI
BshTI (AgeI)
Optimal Buffer: O
A↓CCGGT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R b
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 80°C

Bsp119I

BstBI
Bsp119I (BstBI)
Optimal Buffer: Tango
TT↓CGAA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 20-50
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 80°C

Bsp120I

PspOMI
Bsp120I (PspOMI)
Optimal Buffer: B
G↓GGCCC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R b
1X Tango 100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 80°C

Bsp1407I

BsrGI
Bsp1407I (BsrGI)
Optimal Buffer: Tango
T↓GTACA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

Bsp143I

Sau3AI
Bsp143I (Sau3AI)
Optimal Buffer: Unique
↓GATC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G unique
O onex twox
R b
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

Bsp68I

NruI
Bsp68I (NruI)
Optimal Buffer: O
TCG↓CGA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R b
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.2
Inactivation 65°C

BspLI

NlaIV
BspLI (NlaIV)
Optimal Buffer: Tango
GGN↓NCC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.3
Inactivation 65°C

BspOI

BmtI
BspOI (BmtI)
Optimal Buffer: O
GCTAG↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R b
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 100
Overnight 0.2
Inactivation 80°C

BspPI

AlwI
BspPI (AlwI)
Optimal Buffer: Tango
GGATC(4/5)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R b
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 55°C
Double Digest 0-20
Overnight 0.5
Inactivation 80°C

BspTI

AflII
BspTI (AflII)
Optimal Buffer: O
C↓TTAAG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R b
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 65°C

Bst1107I

BstZ17I
Bst1107I (BstZ17I)
Optimal Buffer: O
GTA↓TAC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R b
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 80°C

BstXI

BstXI
Optimal Buffer: O
CCANNNNN↓NTGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R b
1X Tango 20-50
2X Tango 100
Optimal Temperature 55°C
Double Digest 100
Overnight 0.1
Inactivation 80°C

Bsu15I

ClaI
Bsu15I (ClaI)
Optimal Buffer: Tango
AT↓CGAT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R b
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation 65°C

BsuRI

HaeIII
BsuRI (HaeIII)
Optimal Buffer: R
GG↓CC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R b
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 80°C

BveI

BspMI
BveI (BspMI)
Optimal Buffer: O (+oligo)
ACCTGC(4/8)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X (+oligo)
G orange
O onex twox
R b
1X Tango 0-20 (+oligo)
2X Tango 20-50 (+oligo)
Optimal Temperature 37°C
Double Digest 100 (+oligo)
Overnight 0.2
Inactivation 65°C

CaiI

AlwNI
CaiI (AlwNI)
Optimal Buffer: Tango
CAGNNN↓CTG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R c
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.2
Inactivation 65°C

Cfr10I

BsrFI
Cfr10I (BsrFI)
Optimal Buffer: Unique
R↓CCGGY
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G unique
O onex twox
R c
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation NO

Cfr13I

Sau96I
Cfr13I (Sau96I)
Optimal Buffer: Tango
G↓GNCC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R c
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.3
Inactivation 65°C

Cfr42I

SacII
Cfr42I (SacII)
Optimal Buffer: B
CCGC↓GG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R c
1X Tango 100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

Cfr9I

XmaI
Cfr9I (XmaI)
Optimal Buffer: Unique
C↓CCGGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G unique
O onex
R c
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

CpoI

RsrII
CpoI (RsrII)
Optimal Buffer: Tango
CG↓GWCCG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R c
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.5
Inactivation 65°C

CseI

HgaI
CseI (HgaI)
Optimal Buffer: R
GACGC(5/10)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X* or 2X
G red
O onex twox
R c
1X Tango NR
2X Tango 50-100*
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.5
Inactivation 80°C (10 u)

Csp6I

CviQI
Csp6I (CviQI)
Optimal Buffer: B
G↓TAC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R c
1X Tango 100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

DpnI

DpnI
Optimal Buffer: Tango
Gm6A↓TC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R d
1X Tango 100
2X Tango 100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 80°C

DraI

DraI
Optimal Buffer: Tango
TTT↓AAA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R d
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation 65°C

Eam1104I

EarI
Eam1104I (EarI)
Optimal Buffer: Tango
CTCTTC(1/4)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R e
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 65°C

Eam1105I

AhdI
Eam1105I (AhdI)
Optimal Buffer: Unique
GACNNN↓NNGTC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G unique
O onex twox
R e
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

Ecl136II

EcoICRI
Ecl136II (EcoICRI)
Optimal Buffer: Unique
GAG↓CTC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G unique
O onex
R e
1X Tango 50-100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

Eco105I

SnaBI
Eco105I (SnaBI)
Optimal Buffer: Tango
TAC↓GTA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R e
1X Tango 100*
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 65°C

Eco130I

StyI
Eco130I (StyI)
Optimal Buffer: O
C↓CWWGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R e
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.2
Inactivation 65°C

Eco147I

StuI
Eco147I (StuI)
Optimal Buffer: B
AGG↓CCT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R e
1X Tango 100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation 80°C

Eco24I

BanII
Eco24I (BanII)
Optimal Buffer: Tango
GRGCY↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R e
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

Eco31I

BsaI
Eco31I (BsaI)
Optimal Buffer: G
GGTCTC(1/5)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G green
O onex twox
R e
1X Tango 50-100
2X Tango 100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.3
Inactivation 65°C

Eco32I

EcoRV
Eco32I (EcoRV)
Optimal Buffer: R
GAT↓ATC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R e
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 80°C

Eco47I

AvaII
Eco47I (AvaII)
Optimal Buffer: R
G↓GWCC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R e
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.3
Inactivation 65°C

Eco47III

AfeI
Eco47III (AfeI)
Optimal Buffer: O
AGC↓GCT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R e
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 65°C

Eco52I

EagI
Eco52I (EagI)
Optimal Buffer: Unique
C↓GGCCG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G unique
O twox
R e
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

Eco57I

AcuI
Eco57I (AcuI)
Optimal Buffer: G (+SAM)
CTGAAG(16/14)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X (+SAM)
G green
O onex twox
R e
1X Tango 100 (+SAM)
2X Tango 100 (+SAM)
Optimal Temperature 37°C
Double Digest 20-50 (+SAM)
Overnight 1
Inactivation 65°C

Eco72I

PmlI
Eco72I (PmlI)
Optimal Buffer: Tango
CAC↓GTG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R e
1X Tango NR
2X Tango NR
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 65°C

Eco81I

Bsu36I
Eco81I (Bsu36I)
Optimal Buffer: Tango
CC↓TNAGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R e
1X Tango 50-100
2X Tango 100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 80°C

Eco88I

AvaI
Eco88I (AvaI)
Optimal Buffer: Tango
C↓YCGRG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R e
1X Tango 100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

Eco91I

BstEII
Eco91I (BstEII)
Optimal Buffer: O
G↓GTNACC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R e
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 65°C

EcoO109I

DraII
EcoO109I (DraII)
Optimal Buffer: Tango
RG↓GNCCY
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R e
1X Tango 50-100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.2
Inactivation 65°C

EcoRI

EcoRI
Optimal Buffer: Unique
G↓AATTC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G unique
O twox
R e
1X Tango 0-20
2X Tango NR
Optimal Temperature 37°C
Double Digest 100
Overnight 0.2
Inactivation 65°C

EcoRII

EcoRII
Optimal Buffer: O
↓CCWGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R e
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 80°C

EheI

SfoI
EheI (SfoI)
Optimal Buffer: Tango
GGC↓GCC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R e
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 1
Inactivation 65°C

Esp3I

BsmBI
Esp3I (BsmBI)
Optimal Buffer: Tango (+DTT)
CGTCTC(1/5)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X (+DTT)
G yellow
O onex
R e
1X Tango 100 (+DTT)
2X Tango 20-50 (+DTT)
Optimal Temperature 37°C
Double Digest 0-20 (+DTT)
Overnight 0.2
Inactivation 65°C

FaqI

BsmFI
FaqI (BsmFI)
Optimal Buffer: Tango (+SAM)
GGGAC(10/14)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X (+SAM)
G yellow
O onex twox
R f
1X Tango 20-50 (+SAM)
2X Tango 20-50 (+SAM)
Optimal Temperature 37°C
Double Digest 0-20 (+SAM)
Overnight 1
Inactivation 80°C

FspAI

FspAI
Optimal Buffer: O
RTGC↓GCAY
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R f
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 100
Overnight 0.2
Inactivation 65°C

FspBI

BfaI
FspBI (BfaI)
Optimal Buffer: Tango
C↓TAG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R f
1X Tango 50-100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.3
Inactivation 65°C

GsuI

BpmI
GsuI (BpmI)
Optimal Buffer: B
CTGGAG(16/14)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G blue
O onex twox
R g
1X Tango 100
2X Tango 50-100
Optimal Temperature 30°C
Double Digest 20-50
Overnight 1
Inactivation 65°C

HhaI

HhaI
Optimal Buffer: Tango
GCG↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R h
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation NO

Hin1I

BsaHI
Hin1I (BsaHI)
Optimal Buffer: G
GR↓CGYC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G green
O onex twox
R h
1X Tango 20-50
2X Tango 100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation 65°C

Hin1II

NlaIII
Hin1II (NlaIII)
Optimal Buffer: G
CATG↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G green
O onex twox
R h
1X Tango 50-100
2X Tango 100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.3
Inactivation 65°C

Hin6I

HinP1I
Hin6I (HinP1I)
Optimal Buffer: Tango
G↓CGC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R h
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation 65°C

HincII

HindII
HincII (HindII)
Optimal Buffer: Tango
GTY↓RAC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R h
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation 65°C

HindIII

HindIII
Optimal Buffer: R
A↓AGCTT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R h
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 80°C

HinfI

HinfI
Optimal Buffer: R
G↓ANTC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R h
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 65°C

HpaII

HpaII
Optimal Buffer: Tango
C↓CGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R h
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

HphI

HphI
Optimal Buffer: B
GGTGA(8/7)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R h
1X Tango 100
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

Hpy8I

MjaIV
Hpy8I (MjaIV)
Optimal Buffer: Tango
GTN↓NAC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R h
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 80°C

HpyF10VI

MwoI
HpyF10VI (MwoI)
Optimal Buffer: Tango
GCNNNNN↓NNGC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R h
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 80°C

HpyF3I

DdeI
HpyF3I (DdeI)
Optimal Buffer: Tango
C↓TNAG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R h
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.2
Inactivation 65°C

I-SceI

I-SceI
Optimal Buffer: Tango
TAGGG ATAA↓CAGGGTAAT ATCCC↑TATT GTCCCATTA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R i
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.5
Inactivation 65°C

Kpn2I

BspEI
Kpn2I (BspEI)
Optimal Buffer: Tango
T↓CCGGA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R k
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 55°C
Double Digest 0-20
Overnight 0.2
Inactivation 80°C

KpnI

KpnI
Optimal Buffer: Unique
GGTAC↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G unique
O onex
R k
1X Tango 20-50
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 80°C

KspAI

HpaI
KspAI (HpaI)
Optimal Buffer: B
GTT↓AAC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X* or 2X
G blue
O onex twox
R k
1X Tango 100
2X Tango 50-100*
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.5
Inactivation 65°C

LguI

SapI
LguI (SapI)
Optimal Buffer: Tango
GCTCTTC(1/4)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R l
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.5
Inactivation 65°C

Lsp1109I

BbvI
Lsp1109I (BbvI)
Optimal Buffer: Unique
GCAGC(8/12)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X* or 2X*
G unique
O onex twox
R l
1X Tango 0-20
2X Tango 20-50*
Optimal Temperature 37°C
Double Digest 50-100*
Overnight 0.5
Inactivation 65°C

LweI

SfaNI
LweI (SfaNI)
Optimal Buffer: Tango
GCATC(5/9)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R l
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

MauBI

MauBI
Optimal Buffer: Tango
CG↓CGCGCG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R m
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

MbiI

BsrBI
MbiI (BsrBI)
Optimal Buffer: Tango
CCGCTC(-3/-3)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R m
1X Tango 20-50
2X Tango 100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.2
Inactivation 65°C

MboI

MboI
Optimal Buffer: R
↓GATC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R m
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 65°C

MboII

MboII
Optimal Buffer: B
GAAGA(8/7)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G blue
O onex twox
R m
1X Tango 100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 1
Inactivation 65°C

MlsI

MscI
MlsI (MscI)
Optimal Buffer: R
TGG↓CCA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R m
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 65°C

MluI

MluI
Optimal Buffer: R
A↓CGCGT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R m
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 80°C

MnlI

MnlI
Optimal Buffer: G
CCTC(7/6)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G green
O onex twox
R m
1X Tango 50-100
2X Tango 100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.5
Inactivation 65°C

Mph1103I

NsiI
Mph1103I (NsiI)
Optimal Buffer: R
ATGCA↓T
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R m
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.3
Inactivation 65°C

MreI

Sse232I
MreI (Sse232I)
Optimal Buffer: G
CG↓CCGGCG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G green
O onex
R m
1X Tango 20-50
2X Tango 100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 80°C

MspI

HpaII
MspI (HpaII)
Optimal Buffer: Tango
C↓CGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R m
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.3
Inactivation 80°C

MssI

PmeI
MssI (PmeI)
Optimal Buffer: B
GTTT↓AAAC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R m
1X Tango 100
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 65°C

MunI

MfeI
MunI (MfeI)
Optimal Buffer: G
C↓AATTG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G green
O onex
R m
1X Tango 100
2X Tango 100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

Mva1269I

BsmI
Mva1269I (BsmI)
Optimal Buffer: R
GAATGC(1/-1)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G red
O twox
R m
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 65°C

MvaI

BstNI
MvaI (BstNI)
Optimal Buffer: R
CC↓WGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X* or 2X
G red
O onex twox
R m
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation NO

Nb.Bpu10I

Nb.Bpu10I
Optimal Buffer: R
CCTNA GC
GGANT↑CG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R n
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.3
Inactivation 80°C

Nb.Mva1269I

Nb.Mva1269I
Optimal Buffer: O
GAATG C CTTAC↑G
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R n
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.3
Inactivation 80°C

NcoI

NcoI
Optimal Buffer: Tango
C↓CATGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R n
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation 65°C

NdeI

NdeI
Optimal Buffer: O
CA↓TATG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R n
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 100
Overnight 0.2
Inactivation 65°C

NheI

NheI
Optimal Buffer: Tango
G↓CTAGC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R n
1X Tango 100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

NmuCI

Tsp45I
NmuCI (Tsp45I)
Optimal Buffer: R
↓GTSAC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R n
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 65°C

NotI

NotI
Optimal Buffer: O
GC↓GGCCGC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R n
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 80°C

NsbI

FspI
NsbI (FspI)
Optimal Buffer: Tango
TGC↓GCA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R n
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

Nt.Bpu10I

Nt.Bpu10I
Optimal Buffer: R
CC↓TNAGC GG ANTCG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G red
O twox
R n
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 100
Overnight 0.3
Inactivation 65°C

OliI

AleI
OliI (AleI)
Optimal Buffer: R
CACNN↓NNGTG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G red
O twox
R o
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

PacI

PacI
Optimal Buffer: Unique
TTAAT↓TAA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B NR
G unique
O 0
R p
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.3
Inactivation 65°C

PaeI

SphI
PaeI (SphI)
Optimal Buffer: B
GCATG↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R p
1X Tango 100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

PagI

BspHI
PagI (BspHI)
Optimal Buffer: O
T↓CATGA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B NR
G orange
O 0
R p
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 100
Overnight 0.2
Inactivation 80°C

PasI

PasI
Optimal Buffer: Unique
CC↓CWGGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B NR
G unique
O 0
R p
1X Tango NR
2X Tango NR
Optimal Temperature 55°C
Double Digest NR
Overnight 0.2
Inactivation 80°C

PauI

BssHII
PauI (BssHII)
Optimal Buffer: R
G↓CGCGC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G red
O twox
R p
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 100
Overnight 0.2
Inactivation 80°C

PdiI

NaeI
PdiI (NaeI)
Optimal Buffer: Tango
GCC↓GGC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R p
1X Tango 50-100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 65°C

PdmI

XmnI
PdmI (XmnI)
Optimal Buffer: Tango
GAANN↓NNTTC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R p
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 65°C

PfeI

TfiI
PfeI (TfiI)
Optimal Buffer: O
G↓AWTC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R p
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 65°C

Pfl23II

BsiWI
Pfl23II (BsiWI)
Optimal Buffer: Tango
C↓GTACG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R p
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.3
Inactivation 65°C

PfoI

PfoI
Optimal Buffer: Tango
T↓CCNGGA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R p
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 65°C

Ppu21I

BsaAI
Ppu21I (BsaAI)
Optimal Buffer: Unique
YAC↓GTR
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B NR
G unique
O 0
R p
1X Tango 50-100*
2X Tango 100*
Optimal Temperature 30°C
Double Digest 20-50
Overnight 0.5
Inactivation 65°C

PscI

PciI
PscI (PciI)
Optimal Buffer: Tango
A↓CATGT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R p
1X Tango 20-50
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

Psp1406I

AclI
Psp1406I (AclI)
Optimal Buffer: Tango
AA↓CGTT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R p
1X Tango 100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 65°C

Psp5II

PpuMI
Psp5II (PpuMI)
Optimal Buffer: G
RG↓GWCCY
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G green
O onex twox
R p
1X Tango 0-20
2X Tango 100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.2
Inactivation 80°C

PstI

PstI
Optimal Buffer: O
CTGCA↓G
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R p
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 100
Overnight 0.2
Inactivation NO

PsuI

BstYI
PsuI (BstYI)
Optimal Buffer: B
R↓GATCY
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R p
1X Tango 100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 80°C

PsyI

Tth111I
PsyI (Tth111I)
Optimal Buffer: B
GACN↓NNGTC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R p
1X Tango 100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 80°C

PvuI

PvuI
Optimal Buffer: R
CGAT↓CG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R p
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.2
Inactivation 80°C (10 u)

PvuII

PvuII
Optimal Buffer: G
CAG↓CTG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X* or 2X*
G green
O onex twox
R p
1X Tango 50-100*
2X Tango 100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.2
Inactivation NO

RsaI

RsaI
Optimal Buffer: Tango
GT↓AC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R r
1X Tango 50-100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 80°C

RseI

MslI
RseI (MslI)
Optimal Buffer: R
CAYNN↓NNRTG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R r
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.5
Inactivation 65°C

SacI

SacI
Optimal Buffer: Unique
GAGCT↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G unique
O onex twox
R s
1X Tango 50-100
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

SalI

SalI
Optimal Buffer: O
G↓TCGAC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R s
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 65°C

SatI

Fnu4HI
SatI (Fnu4HI)
Optimal Buffer: G
GC↓NGC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G green
O onex twox
R s
1X Tango 20-50
2X Tango 100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation 65°C

ScaI

ScaI
Optimal Buffer: Unique
AGT↓ACT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B NR
G unique
O 0
R s
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.5
Inactivation 80°C (10 u)

SchI

MlyI
SchI (MlyI)
Optimal Buffer: Tango
GAGTC(5/5)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R s
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

SdaI

SbfI
SdaI (SbfI)
Optimal Buffer: Unique
CCTGCA↓GG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G unique
O twox
R s
1X Tango NR
2X Tango NR
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.3
Inactivation 80°C

SduI

Bsp1286I
SduI (Bsp1286I)
Optimal Buffer: Unique
GDGCH↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B NR
G unique
O 0
R s
1X Tango NR
2X Tango 50-100*
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.3
Inactivation 65°C

SfaAI

AsiSI
SfaAI (AsiSI)
Optimal Buffer: Tango
GCGAT↓CGC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R s
1X Tango 50-100
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 80°C

SfiI

SfiI
Optimal Buffer: G
GGCCNNNN↓NGGCC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G green
O onex
R s
1X Tango 50-100
2X Tango 100
Optimal Temperature 50°C
Double Digest 20-50
Overnight 0.2
Inactivation NO

SgeI

SgeI
Optimal Buffer: Unique
m5CNNG(9/13)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B NR
G unique
O 0
R s
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0
Inactivation 65°C

SgrDI

SgrDI
Optimal Buffer: R
CG↓TCGACG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G red
O twox
R s
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.3
Inactivation 65°C

SgsI

AscI
SgsI (AscI)
Optimal Buffer: Tango
GG↓CGCGCC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R s
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

SmaI

SmaI
Optimal Buffer: Tango
CCC↓GGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R s
1X Tango 50-100
2X Tango 0-20
Optimal Temperature 30°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

SmiI

SwaI
SmiI (SwaI)
Optimal Buffer: O
ATTT↓AAAT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R s
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 30°C
Double Digest 100
Overnight 0.1
Inactivation 65°C

SmoI

SmlI
SmoI (SmlI)
Optimal Buffer: Tango
C↓TYRAG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R s
1X Tango 50-100
2X Tango 20-50
Optimal Temperature 55°C
Double Digest 0-20
Overnight 0.2
Inactivation 80°C

SsiI

AciI
SsiI (AciI)
Optimal Buffer: O
CCGC(-3/-1)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 2X
G orange
O twox
R s
1X Tango NR
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 100
Overnight 0.5
Inactivation 65°C

SspDI

KasI
SspDI (KasI)
Optimal Buffer: Tango
G↓GCGCC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R s
1X Tango 20-50
2X Tango 0-20
Optimal Temperature 37°C
Double Digest NR
Overnight 0.1
Inactivation 80°C

SspI

SspI
Optimal Buffer: G
AAT↓ATT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G green
O onex twox
R s
1X Tango 20-50
2X Tango 100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

TaaI

HpyCH4III
TaaI (HpyCH4III)
Optimal Buffer: Tango
ACN↓GT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R t
1X Tango 0-20
2X Tango 0-20
Optimal Temperature 65°C
Double Digest 0-20
Overnight 0.2
Inactivation NO

TaiI

MaeII
TaiI (MaeII)
Optimal Buffer: R
ACGT↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R t
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 65°C
Double Digest 20-50
Overnight 0.3
Inactivation NO

TaqI

TaqI
Optimal Buffer: Unique
T↓CGA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G unique
O onex twox
R t
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 65°C
Double Digest 20-50
Overnight 0.3
Inactivation NO

TasI

Tsp509I
TasI (Tsp509I)
Optimal Buffer: B
↓AATT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R t
1X Tango 100
2X Tango 50-100
Optimal Temperature 65°C
Double Digest 20-50
Overnight 0.3
Inactivation NO

TatI

TatI
Optimal Buffer: Tango
W↓GTACW
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X*
G yellow
O onex
R t
1X Tango NR
2X Tango 50-100*
Optimal Temperature 65°C
Double Digest 20-50
Overnight 0.2
Inactivation NO

TauI

TauI
Optimal Buffer: B
GCSG↓C
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G blue
O onex
R t
1X Tango 100
2X Tango 50-100
Optimal Temperature 55°C
Double Digest 0-20
Overnight 1
Inactivation NO

Tru1I

MseI
Tru1I (MseI)
Optimal Buffer: R
T↓TAA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R t
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 65°C
Double Digest 20-50
Overnight 0.2
Inactivation NO

TscAI

TspRI
TscAI (TspRI)
Optimal Buffer: Tango
CASTG(2/-7)↓
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R t
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 65°C
Double Digest 20-50
Overnight 0.2
Inactivation NO

Van91I

PflMI
Van91I (PflMI)
Optimal Buffer: R
CCANNNN↓NTGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R v
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 65°C

VspI

AseI
VspI (AseI)
Optimal Buffer: O
AT↓TAAT
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G orange
O onex twox
R v
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 100
Overnight 0.1
Inactivation 65°C

XagI

EcoNI
XagI (EcoNI)
Optimal Buffer: R
CCTNN↓NNNAGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R x
1X Tango 0-20
2X Tango 20-50
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 65°C

XapI

ApoI
XapI (ApoI)
Optimal Buffer: Tango
R↓AATTY
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R x
1X Tango 50-100
2X Tango 100
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 80°C

XbaI

XbaI
Optimal Buffer: Tango
T↓CTAGA
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R x
1X Tango 50-100
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 20-50
Overnight 0.1
Inactivation 65°C

XceI

NspI
XceI (NspI)
Optimal Buffer: Tango
RCATG↓Y
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X
G yellow
O onex
R x
1X Tango 50-100
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.2
Inactivation 65°C

XhoI

XhoI
Optimal Buffer: R
C↓TCGAG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G red
O onex twox
R x
1X Tango 0-20
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.1
Inactivation 80°C

XmaJI

AvrII
XmaJI (AvrII)
Optimal Buffer: Tango
C↓CTAGG
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G yellow
O onex twox
R x
1X Tango 20-50
2X Tango 50-100
Optimal Temperature 37°C
Double Digest 50-100
Overnight 0.2
Inactivation 80°C

XmiI

AccI
XmiI (AccI)
Optimal Buffer: B
GT↓MKAC
  • Conditions
  • Methylation
  • Efficiency
  • Recognition Sites
B 1X or 2X
G blue
O onex twox
R x
1X Tango 100
2X Tango 0-20
Optimal Temperature 37°C
Double Digest 0-20
Overnight 0.1
Inactivation 65°C

Notes

* Star activity appears at a geater than 5-fold overdigestion (5 units x 1 hour).

NR - buffer is not recommended, because of high star activity.

80°C(10U)- only small amounts of the restriction enzyme (up to 10 units) can be inactivated at 80°C in 20 minutes.