Anza restriction enzyme selection tool search page

Use our restriction site finder tool to find the Anza restriction enzymes that best suit your research needs. Search by product name, isoschizomer name, recognition sequence, or SKU number. Search results include restriction enzyme names, cut site, overhang, compatible ends, isoschizomers, and enzyme properties.

Have questions about how restriction enzymes work, where they get their names, or how to troubleshoot common problems? Explore our Restriction Enzyme education resources in the Invitrogen School of Molecular Biology.

Learn more about the Anza Restriction Enzyme Cloning System


B = C or G or T
D = A or G or T
H = A or C or T
K = G or T
M = A or C
N = A or C or G or T

R = A or G
S = C or G
V = A or C or G
W = A or T
Y = C or T

Product name,
Cat. No.
Iso-
schizomers
Sequence and cut siteOverhangAnza restriction enzymes resulting in compatible endsProperties
Anza 1 NotI,
IVGN0014
CciNIGCGGCCGC

5'...G C^G G C C G C...3'
3'...C G C C G G^C G...5'
5’ protruding endsAnza 47 Eco52I, Anza 121 Bsp120IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 2 NcoI,
IVGN0026
Bsp19ICCATGG

5'...C^C A T G G...3'
3'...G G T A C^C...5'
5’ protruding endsAnza 58 PagIAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 3 BcuI,
IVGN0034
AhlI, SpeIACTAGT

5'...A^C T A G T...3'
3'...T G A T C^A...5'
5’ protruding endsAnza 6 NheI, Anza 12 XbaI, Anza 15 XmaJIAnza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 4 BpiI,
IVGN0044
BbsI, BstV2IGAAGAC(2/6)

5'...G A A G A C (N)2^...3'
3'...C T T C T G (N)6^...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 5 BamHI,
IVGN0056
GGATCC

5'...G^G A T C C...3'
3'...C C T A G^G...5'
5’ protruding endsAnza 19 BglII, Anza 55 MboI, Anza 73 BclI, Anza 97 Bsp143IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 6 NheI,
IVGN0066
AsuNHIGCTAGC

5'...G^C T A G C...3'
3'...C G A T C^G...5'
5’ protruding endsAnza 3 BcuI, Anza 12 XbaI, Anza 15 XmaJIAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 7 BshTI,
IVGN0074
AsiGI, CspAI, PinAI, AgeIACCGGT

5'...A^C C G G T...3'
3'...T G G C C^A...5'
5’ protruding endsAnza 60 Kpn2I, Anza 85 MreI, Anza 90 Eco88I* (*if recognized sequence is 5'-CCCGGG-3')Anza universal protocol
no heat inactivation
2-step, column purify first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 8 XhoI,
IVGN0086
PaeR7I, Sfr274I, SlaICTCGAG

5'...C^T C G A G...3'
3'...G A G C T^C...5'
5’ protruding endsAnza 14 SalI, Anza 90 Eco88I* (*if recognized sequence is 5'-CTCGAG-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 9 NdeI,
IVGN0096
FauNDICATATG

5'...C A^T A T G...3'
3'...G T A T^A C...5'
5’ protruding endsAnza 64 SaqAI, Anza 76 VspI, Anza 80 FspBI, Anza 122 Csp6IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 10 DpnI,
IVGN0106
MalIGm6ATC

5'...G m6A^T C...3'
3'...C T^m6A G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
Anza 11 EcoRI,
IVGN0116
GAATTC

5'...G^A A T T C...3'
3'...C T T A A^G...5'
5’ protruding endsAnza 31 MunI, Anza 111 XapIAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 12 XbaI,
IVGN0126
TCTAGA

5'...T^C T A G A...3'
3'...A G A T C^T...5'
5’ protruding endsAnza 3 BcuI, Anza 6 NheI, Anza 15 XmaJIAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping Dam methylation which will result in blocked or impaired DNA cleavage
Anza 13 Esp3I,
IVGN0136
BsmBI CGTCTC(1/5)

5'...C G T C T C (N)1^...3'
3'...G C A G A G (N)5^...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 14 SalI,
IVGN0146
GTCGAC

5'...G^T C G A C...3'
3'...C A G C T^G...5'
5’ protruding endsAnza 8 XhoI, Anza 90 Eco88I* (*if recognized sequence is 5'-CTCGAG-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza15 XmaJI,
IVGN0154
AspA2I, AvrII, BlnI CCTAGG

5'...C^C T A G G...3'
3'...G G A T C^C...5'
5’ protruding endsAnza 3 BcuI, Anza 6 NheI, Anza 12 XbaIAnza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 16 HindIII,
IVGN0166
AAGCTT

5'...A^A G C T T...3'
3'...T T C G A^A...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 17 KpnI,
IVGN0176
GGTACC

5'...G G T A C^C...3'
3'...C^C A T G G...5'
3’ protruding endsAnza universal protocol
no heat inactivation
2-step, column purify first
Anza 18 PacI,
IVGN0184
TTAATTAA

5'…T T A A T^T A A...3'
3'…A A T^T A A T T...5'
3’ protruding endsAnza 27 PvuI, Anza 40 SfaAIAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 19 BglII,
IVGN0196
AGATCT

5'...A^G A T C T...3'
3'...T C T A G^A...5'
5’ protruding endsAnza 5 BamHI, Anza 55 MboI, Anza 73 BclI, Anza 97 Bsp143IAnza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 20 SacI,
IVGN0206
Psp124BI, SstIGAGCTC

5'...G A G C T^C...3'
3'...C^T C G A G...5'
3’ protruding endsAnza 126 Alw21I* (*if recognized sequence is 5'-GAGCTC-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 21 SgsI,
IVGN0214
AscI, PalAI GGCGCGCC

5'...G G^C G C G C C...3'
3'...C C G C G C^G G...5'
5’ protruding endsAnza 28 MluI, Anza 45 PteI, Anza 95 MauBIAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 22 SmaI,
IVGN0226
CCCGGG

5'...C C C^G G G...3'
3'...G G G^C C C...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 23 PstI,
IVGN0236
CTGCAG

5'...C T G C A^G...3'
3'...G^A C G T C...5'
3’ protruding endsAnza 37 Mph1103I, Anza 126 Alw21I* (*if recognized sequence is 5'-GTGCAC-3')Anza universal protocol
no heat inactivation
2-step, column purify first
Anza 24 MssI,
IVGN0244
PmeIGTTTAAAC

5'...G T T T^A A A C...3'
3'...C A A A^T T T G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 25 PaeI,
IVGN0254
SphI GCATGC

5'...G C A T G^C...3'
3'...C^G T A C G...5'
3’ protruding endsAnza 56 Hin1II, Anza 98 XceIAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 26 Eco32I,
IVGN0266
EcoRV GATATC

5'...G A T^A T C...3'
3'...C T A^T A G...5'
bluntAnza universal protocol
no heat inactivation
2-step, column purify first
Anza 27 PvuI,
IVGN0274
Ple19I CGATCG

5'...C G A T^C G...3'
3'...G C^T A G C...5'
3’ protruding endsAnza 18 PacI, Anza 40 SfaAIAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 28 MluI,
IVGN0286
ACGCGT

5'...A^C G C G T...3'
3'...T G C G C^A...5'
5’ protruding endsAnza 21 SgsI, Anza 45 PteI, Anza 95 MauBIAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 29 KflI,
IVGN0294
GGGWCCC

5'…G G^G W C C C...3'
3'…C C C W G^G G...5'
5’ protruding endsAnza 63 CpoI, Anza 87 Eco47IAnza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 30 Bsu15I,
IVGN0306
Bsa29I, BseCI, BshVI, BspDI, BsuTUI, ClaIATCGAT

5'...A T^C G A T...3'
3'...T A G C^T A...5'
5’ protruding endsAnza 65 MspI, Anza 88 Bsp119I, Anza 93 HpaII, Anza 103 Psp1406I, Anza 105 Hin1I, Anza 123 Hin6I, Anza 96 XmiI* (*if recognized sequence is 5'-GTCGAC-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequenceThe target site may have overlapping Dam methylation which will result in blocked or impaired DNA cleavage
Anza 31 MunI,
IVGN0314
MfeI CAATTG

5'...C^A A T T G...3'
3'...G T T A A^C...5'
5’ protruding endsAnza 11 EcoRI, Anza 111 XapIAnza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 32 ApaI,
IVGN0326
GGGCCC

5'...G G G C C^C...3'
3'...C^C C G G G...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavageThe target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 33 LguI,
IVGN0334
BspQI, PciSI, SapI GCTCTTC(1/4)

5'...G C T C T T C (N)1^...3'
3'...C G A G A A G (N)4^...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 34 Pfl23II,
IVGN0344
BsiWI, PspLI CGTACG

5'...C^G T A C G...3'
3'...G C A T G^C...5'
5’ protruding endsAnza 39 Bsp1407I, Anza 91 Acc65I, Anza 129 BshNI* (*if recognized sequence is 5'-GGTACC-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 35 Eco47III,
IVGN0354
AfeI, Aor51HI AGCGCT

5'...A G C^G C T...3'
3'...T C G^C G A...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 36 Eco31I,
IVGN0366
BsaI, Bso31I, BspTNIGGTCTC(1/5)

5'...G G T C T C (N)1^...3'
3'...C C A G A G (N)5^...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavageThe target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 37 Mph1103I,
IVGN0376
EcoT22I, NsiI, Zsp2I ATGCAT

5'...A T G C A^T...3'
3'...T^A C G T A...5'
3’ protruding endsAnza 23 PstI, Anza 126 Alw21I* (*if recognized sequence is 5'-GTGCAC-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 38 ScaI,
IVGN0386
BmcAI, ZrmI AGTACT

5'...A G T^A C T...3'
3'...T C A^T G A...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 39 Bsp1407I,
IVGN0396
BsrGI, BstAUI TGTACA

5'...T^G T A C A...3'
3'...A C A T G^T...5'
5’ protruding endsAnza 34 Pfl23II, Anza 91 Acc65I, Anza 129 BshNI* (*if recognized sequence is 5'-GGTACC-3')Anza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 40 SfaAI,
IVGN0404
AsiSI, RgaI, SgfI GCGATCGC

5'...G C G A T^C G C...3'
3'...C G C^T A G C G...5'
3’ protruding endsAnza 18 PacI, Anza 27 PvuIAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 41 HpyF3I,
IVGN0416
BstDEI, DdeI CTNAG

5'...C^T N A G...3'
3'...G A N T^C...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 42 RsaI,
IVGN0426
AfaIGTAC

5'...G T^A C...3'
3'...C A^T G...5'
bluntAnza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 43 Eco105I,
IVGN0434
BstSNI, SnaBI TACGTA

5'...T A C^G T A...3'
3'...A T G^C A T...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 44 AluI,
IVGN0446
AluBI AGCT

5'...A G^C T...3'
3'...T C^G A...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 45 PteI,
IVGN0454
BsePI, BssHII, PauIGCGCGC

5'...G^C G C G C...3'
3'...C G C G C^G...5'
5’ protruding endsAnza 21 SgsI, Anza 28 MluI, Anza 95 MauBIAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 46 AatII,
IVGN0464
GACGTC

5'...G A C G T^C...3'
3'...C^T G C A G...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 47 Eco52I,
IVGN0474
BseX3I, BstZI, EagI, EclXI CGGCCG

5'...C^G G C C G...3'
3'...G C C G G^C...5'
5’ protruding endsAnza 1 NotI, Anza 121 Bsp120IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 48 MnlI,
IVGN0484
CCTC(7/6)

5'...C C T C (N)7^...3'
3'...G G A G (N)6^...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 49 SmiI,
IVGN0496
SwaI ATTTAAAT

5'...A T T T^A A A T...3'
3'...T A A A^T T T A...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 50 KspAI,
IVGN0504
HpaI GTTAAC

5'...G T T^A A C...3'
3'...C A A^T T G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 51 BspTI,
IVGN0516
AflII, BfrI, BstAFI, MspCI, Vha464I CTTAAG

5'...C^T T A A G...3'
3'...G A A T T^C...5'
5’ protruding endsAnza universal protocol
no heat inactivation
2-step, column purify first
Anza 52 PvuII,
IVGN0526
CAGCTG

5'...C A G^C T G...3'
3'...G T C^G A C...5'
bluntAnza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 53 AanI,
IVGN0534
PsiI TTATAA

5'...T T A^T A A...3'
3'...A A T^A T T...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 54 Eco147I,
IVGN0546
PceI, SseBI, StuI AGGCCT

5'...A G G^C C T...3'
3'...T C C^G G A...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 55 MboI,
IVGN0554
BfuCI, Bsp143I, BssMI, BstMBI, DpnII, Kzo9I, NdeII, Sau3AIGATC

5'...^G A T C ...3'
3'... C T A G^...5'
5’ protruding endsAnza 5 BamHI, Anza 19 BglII, Anza 73 BclI, Anza 97 Bsp143IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by Dam methylation within the target sequenc
Anza 56 Hin1II,
IVGN0566
FaeI, Hsp92II, NlaIII CATG

5'... C A T G^...3'
3'...^G T A C ...5'
3’ protruding endsAnza 25 PaeI, Anza 98 XceIAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 57 Bpu1102I,
IVGN0574
BlpI, Bsp1720I GCTNAGC

5'...G C^T N A G C...3'
3'...C G A N T^C G...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
Anza 58 PagI,
IVGN0584
CciI, BspHI TCATGA

5'...T^C A T G A...3'
3'...A G T A C^T...5'
5’ protruding endsAnza 2 NcoIAnza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping Dam methylation which will result in blocked or impaired DNA cleavage
Anza 59 HhaI,
IVGN0596
AspLEI, BstHHI, CfoI GCGC

5'...G C G^C...3'
3'...C^G C G...5'
3’ protruding endsAnza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 60 Kpn2I,
IVGN0606
AccIII, Aor13HI, BseAI, Bsp13I, BspEI, MroI TCCGGA

5'...T^C C G G A...3'
3'...A G G C C^T...5'
5’ protruding endsAnza 7 BshTI, Anza 85 MreI, Anza 90 Eco88I* (*if recognized sequence is 5'-CCCGGG-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 61 PfoI,
IVGN0614
TCCNGGA

5'...T^C C N G G A...3'
3'...A G G N C C^T...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavageThe target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavageThe target site may have overlapping Dam methylation which will result in blocked or impaired DNA cleavage
Anza 62 MlsI,
IVGN0624
BalI, MluNI, MscI, Msp20I, BspTNITGGCCA

5'...T G G^C C A...3'
3'...A C C^G G T...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 63 CpoI,
IVGN0634
CspI, Rsr2I, RsrII CGGWCCG

5'...C G^G W C C G...3'
3'...G C C W G^G C...5'
5’ protruding endsAnza 28 KflI, Anza 87 Eco47IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 64 SaqAI,
IVGN0644
MseI, Tru1I, Tru9ITTAA

5'...T^T A A...3'
3'...A A T^T...5'
5’ protruding endsAnza 9 NdeI, Anza 76 VspI, Anza 80 FspBI, Anza 122 Csp6IAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 65 MspI,
IVGN0656
BsiSI, HapII, Hpall CCGG

5'...C^C G G...3'
3'...G G C^C...5'
5’ protruding endsAnza 30 Bsu15I, Anza 88 Bsp119I, Anza 93 HpaII, Anza 103 Psp1406I, Anza 105 Hin1I, Anza 123 Hin6I, Anza 96 XmiI* (*if recognized sequence is 5'-GTCGAC-3')Anza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 66 BstXI,
IVGN0666
CCANNNNNNTGG

5'...C C A N N N N N^N T G G...3'
3'...G G T N^N N N N N A C C...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
The target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 67 RruI,
IVGN0676
Bsp68I, BtuMI, NruITCGCGA

5'...T C G^C G A...3'
3'...A G C^G C T...5'
bluntAnza universal protocol
no heat inactivation
2-step, column purify first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 68 BsuRI,
IVGN0686
BshFI, BsnI, HaeIII, BspANIGGCC

5'...G G^C C...3'
3'...C C^G G...5'
bluntAnza universal protocol
no heat inactivation
2-step, column purify first
Anza 69 BglI,
IVGN0696
GCCNNNNNGGC

5'...G C C N N N N^N G G C...3'
3'...C G G N^N N N N C C G...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 70 NsbI,
IVGN0704
Acc16I, FspITGCGCA

5'...T G C^G C A...3'
3'...A C G^C G T...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 71 HinfI,
IVGN0716
GANTC

5'...G^A N T C...3'
3'...C T N A^G...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 72 HincII,
IVGN0726
HindII GTYRAC

5'...G T Y^R A C...3'
3'...C A R^Y T G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 73 BclI,
IVGN0736
FbaI, Ksp22I TGATCA

5'...T^G A T C A...3'
3'...A C T A G^T...5'
5’ protruding endsAnza 5 BamHI, Anza 19 BglII, Anza 55 MboI, Anza 97 Bsp143IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by Dam methylation within the target sequenc
Anza 74 CsiI,
IVGN0744
ACCWGGT

5'…A^C C W G G T...3'
3'…T G G W C C^A...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 75 Alw44I,
IVGN0756
ApaLI, VneI GTGCAC

5'...G^T G C A C...3'
3'...C A C G T^G...5'
5’ protruding endsAnza 94 BfmI* (*if recognized sequence is 5'-CTGCAG-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 76 VspI,
IVGN0766
AseI, PshBI ATTAAT

5'...A T^T A A T...3'
3'...T A A T^T A...5'
5’ protruding endsAnza 9 NdeI, Anza 64 SaqAI, Anza 80 FspBI, Anza 122 Csp6IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 77 DraI,
IVGN0776
TTTAAA

5'...T T T^A A A...3'
3'...A A A^T T T...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
Anza 78 AdeI,
IVGN0786
DraIII CACNNNGTG

5'...C A C N N N^G T G...3'
3'...G T G^N N N C A C...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 79 PdiI,
IVGN0794
NaeIGCCGGC

5'...G C C^G G C...3'
3'...C G G^C C G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 80 FspBI,
IVGN0804
BfaI, MaeI, XspI, SspMl CTAG

5'...C^T A G...3'
3'...G A T^C...5'
5’ protruding endsAnza 9 NdeI, Anza 64 SaqAI, Anza 76 VspI, Anza 122 Csp6IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 81 Eco91I,
IVGN0816
BstEII, BstPI, EcoO65I, PspEI GGTNACC

5'...G^G T N A C C...3'
3'...C C A N T G^G...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
Anza 82 Eco72I,
IVGN0826
AcvI, BbrPI, PmaCI, Pm1I, PspCI CACGTG

5'...C A C^G T G...3'
3'...G T G^C A C...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 83 Eco81I,
IVGN0836
AxyI, Bse21I, Bsu36I CCTNAGG

5'...C C^T N A G G...3'
3'...G G A N T^C C...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 84 FspAI,
IVGN0844
RTGCGCAY

5'...R T G C^G C A Y...3'
3'...Y A C G^C G T R...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 85 MreI,
IVGN0854
CGCCGGCG

5'...C G^C C G G C G...3'
3'...G C G G C C^G C...5'
5’ protruding endsAnza 7 BshTI, Anza 60 Kpn2I, Anza 90 Eco88I* (*if recognized sequence is 5'-CCCGGG-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 86 PdmI,
IVGN0866
Asp700I, MroXI, XmnI GAANNNNTTC

5'...G A A N N^N N T T C...3'
3'...C T T N N^N N A A G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 87 Eco47I,
IVGN0876
AvaII, Bme18I, SinI, VpaK11BI GGWCC

5'...G^G W C C...3'
3'...C C W G^G...5'
5’ protruding endsAnza 28, KflI, Anza 63 CpoIAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavageThe target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 88 Bsp119I,
IVGN0886
AsuII, Bpu14I, BspT104I, BstBI, NspV, SfuITTCGAA

5'...T T^C G A A...3'
3'...A A G C^T T...5'
5’ protruding endsAnza 30 Bsu15I, Anza 65 MspI, Anza 93 HpaII, Anza 103 Psp1406I, Anza 105 Hin1I, Anza 123 Hin6I, Anza 96 XmiI* (*if recognized sequence is 5'-GTCGAC-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 89 Mva1269I,
IVGN0894
BsmI, PctI GAATGC(1/-1)

5'...G A A T G C N^...3'
3'...C T T A C^G N ...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 90 Eco88I,
IVGN0906
Ama87I, AvaI, BsiHKCI, BsoBI, BmeT110lCYCGRG

5'...C^Y C G R G...3'
3'...G R G C Y^C...5'
5’ protruding endsIf recognized sequence is 5'-CTCGAG-3', then generate compatible ends for Anza 8 XhoI, Anza 14 SalI. If recognized sequence is 5'-CCCGGG-3', then generate compatible ends for Anza 7 BshTI, Anza 60 Kpn2I, Anza 85 MreIAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 91 Acc65I,
IVGN0916
Asp718IGGTACC

5'...G^G T A C C...3'
3'...C C A T G^G...5'
5’ protruding endsAnza 34 Pfl23II, Anza 39 Bsp1407I, Anza 129 BshNI* (*if recognized sequence is 5'-GGTACC-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavageThe target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 92 EheI,
IVGN0924
DinI, EgeI, SfoIGGCGCC

5'...G G C^G C C...3'
3'...C C G^C G G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 93 HpaII,
IVGN0936
BsiSI, HapII, MsplCCGG

5'...C^C G G...3'
3'...G G C^C...5'
5’ protruding endsAnza 30 Bsu15I, Anza 65 MspI, Anza 88 Bsp119I, Anza 103 Psp1406I, Anza 105 Hin1I, Anza 123 Hin6I, Anza 96 XmiI* (*if recognized sequence is 5'-GTCGAC-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 94 BfmI,
IVGN0944
BstSFI, SfcI CTRYAG

5'...C^T R Y A G...3'
3'...G A Y R T^C...5'
5’ protruding endsIf recognized sequnce is 5'-CTGCAG-3', then generate compatible ends for Anza 75 Alw44IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 95 MauBI,
IVGN0954
CGCGCGCG

5'...C G^C G C G C G...3'
3'...G C G C G C^G C...5'
5’ protruding endsAnza 21 SgsI, Anza 28 MluI, Anza 45 PteIAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 96 XmiI,
IVGN0966
AccI, FblI GTMKAC

5'...G T^M K A C...3'
3'...C A K M^T G...5'
5’ protruding ends If recognized sequence is 5'-GTCGAC-3', then generate compatible ends for Anza 30 Bsu15I, Anza 65 MspI, Anza 88 Bsp119I, Anza 93 HpaII, Anza 103 Psp1406I, Anza 105 Hin1I, Anza 123 Hin6IAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 97 Bsp143I,
IVGN0976
BfuCI, BssMI, BstMBI, DpnII, Kzo9I, MboI, NdeII, Sau3AIGATC

5'...^G A T C ...3'
3'... C T A G^...5'
5’ protruding endsAnza 5 BamHI, Anza 19 BglII, Anza 55 MboI, Anza 73 BclIAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 98 XceI,
IVGN0984
BstNSI, NspI RCATGY

5'...R C A T G^Y...3'
3'...Y^G T A C R...5'
3’ protruding endsAnza 25 PaeI, Anza 56 Hin1IIAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 99 XagI,
IVGN0996
BstENI, EcoNI CCTNNNNNAGG

5'...C C T N N^N N N A G G...3'
3'...G G A N N N^N N T C C...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
Anza 100 Bsh1236I,
IVGN1006
AccII, BspFNI, BstFNI, BstUI, MvnI CGCG

5'...C G^C G...3'
3'...G C^G C...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 101 BoxI,
IVGN1016
BstPAI, PshAI GACNNNNGTC

5'...G A C N N^N N G T C...3'
3'...C T G N N^N N C A G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 102 CaiI,
IVGN1024
AlwNI, PstNICAGNNNCTG

5'...C A G N N N^C T G...3'
3'...G T C^N N N G A C...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
The target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 103 Psp1406I,
IVGN1034
AclI AACGTT

5'...A A^C G T T...3'
3'...T T G C^A A...5'
5’ protruding endsAnza 30 Bsu15I, Anza 65 MspI, Anza 88 Bsp119I, Anza 93 HpaII, Anza 105 Hin1I, Anza 123 Hin6I, Anza 96 XmiI* (*if recognized sequence is 5'-GTCGAC-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 104 MboII,
IVGN1044
GAAGA(8/7)

5'...G A A G A (N)8^...3'
3'...C T T C T (N)7^...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping Dam methylation which will result in blocked or impaired DNA cleavage
Anza 105 Hin1I,
IVGN1056
AcyI, BsaHI, BssNI, BstACI, Hsp92I GRCGYC

5'...G R^C G Y C...3'
3'...C Y G C^R G...5'
5’ protruding endsAnza 30 Bsu15I, Anza 65 MspI, Anza 88 Bsp119I, Anza 93 HpaII, Anza 103 Psp1406I, Anza 123 Hin6I, Anza 96 XmiI* (*if recognized sequence is 5'-GTCGAC-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequenceThe target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 106 Van91I,
IVGN1066
AccB7I, PflMICCANNNNNTGG

5'...C C A N N N N^N T G G...3'
3'...G G T N^N N N N A C C...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
The target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 107 BspLI,
IVGN1074
BmiI, NlaIV, PspN4I GGNNCC

5'...G G N^N C C...3'
3'...C C N^N G G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavageThe target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 108 SatI,
IVGN1084
Fnu4HI, Fsp4HI GCNGC

5'...G C^N G C...3'
3'...C G N^C G...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 109 Alw26I,
IVGN1096
BcoDI, BsmAI, BstMAI GTCTC(1/5)

5'...G T C T C (N)1^...3'
3'...C A G A G (N)5^...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 111 XapI,
IVGN1116
AcsI, ApoI RAATTY

5'...R^A A T T Y...3'
3'...Y T T A A^R...5'
5’ protruding endsAnza 11 EcoRI, Anza 31 MunIAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
Anza 112 BseGI,
IVGN1126
BstF5I, BtsCI GGATG(2/0)

5'...G G A T G N N^...3'
3'...C C T A C^N N ...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
Anza 113 BcnI,
IVGN1136
AsuC2I, BpuMI, NciI CCSGG

5'...C C^S G G...3'
3'...G G S^C C...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 114 Hpy8I,
IVGN1146
Hpy166II GTNNAC

5'...G T N^N A C...3'
3'...C A N^N T G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 115 MbiI,
IVGN1156
AccBSI, BsrBI CCGCTC (-3/-3)

5'...C C G^C T C...3'
3'...G G C^G A G...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 116 Cfr13I,
IVGN1166
AspS9I, BmgT120I, PspPI, Sau96I GGNCC

5'...G^G N C C...3'
3'...C C N G^G...5'
5’ protruding endsAnza universal protocol
no heat inactivation
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavageThe target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 117 EcoO109I,
IVGN1176
RGGNCCY

5'...R G^G N C C Y...3'
3'...Y C C N G^G R...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
The target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 118 BseDI,
IVGN1186
BsaJI, BssECI CCNNGG

5'...C^C N N G G...3'
3'...G G N N C^C...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 119 BmsI,
IVGN1194
LweI, SfaNIGCATC (5/9)

5'…G C A T C (N)5^...3'
3'…C G T A G (N)9^...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 120 NmuCI,
IVGN1204
TseFI, Tsp45I GTSAC

5'...^G T S A C ...3'
3'... C A S T G^...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 121 Bsp120I,
IVGN1216
PspOMI GGGCCC

5'...G^G G C C C...3'
3'...C C C G G^G...5'
5’ protruding endsAnza 1 NotI, Anza 47 Eco52IAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavageThe target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage
Anza 122 Csp6I,
IVGN1226
CviQI, RsaNIGTAC

5'...G^T A C...3'
3'...C A T^G...5'
5’ protruding endsAnza 9 NdeI, Anza 64 SaqAI, Anza 76 VspI, Anza 80 FspBIAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 123 Hin6I,
IVGN1236
HinP1I, HspAI GCGC

5'...G^C G C...3'
3'...C G C^G...5'
5’ protruding endsAnza 30 Bsu15I, Anza 65 MspI, Anza 88 Bsp119I, Anza 93 HpaII, Anza 103 Psp1406I, Anza 105 Hin1I, Anza 96 XmiI* (*if recognized sequence is 5'-GTCGAC-3')Anza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
DNA cleavage by the restriction enzyme is blocked or impaired by CpG methylation within the target sequence
Anza 124 PfeI,
IVGN1244
TfiI GAWTC

5'...G^A W T C...3'
3'...C T W A^G...5'
5’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 125 HpyF10VI,
IVGN1254
BstMWI, MwoIGCNNNNNNNGC

5'...G C N N N N N^N N G C...3'
3'...C G N N^N N N N N C G...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 126 Alw21I,
IVGN1266
Bbv12I, BsiHKAI GWGCWC

5'...G W G C W^C...3'
3'...C^W C G W G...5'
3’ protruding endsIf recognized sequence is 5'-GAGCTC-3', then generate compatible ends for Anza 20 SacI. If recognized sequence is 5'-GTGCAC-3', then generate compatible ends for Anza 23 PstI, Anza 37 Mph1103IAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
Anza 127 RseI,
IVGN1274
MslI, SmiMI CAYNNNNRTG

5'...C A Y N N^N N R T G...3'
3'...G T R N N^N N Y A C...5'
bluntAnza universal protocol
heat inactivate at 80°C for 20 minutes
1-step restriction enzyme digestion and alkaline phosphatase treatment
Anza 128 PspFI,
IVGN1284
GsaICCCAGC(-1/-5)

5'…C C C A G^C...3'
3'…G^G G T C G...5'
3’ protruding endsAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, column purify first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavage
Anza 129 BshNI,
IVGN1294
AccB1I, BanI, BspT107I GGYRCC

5'...G^G Y R C C...3'
3'...C C R Y G^G...5'
5’ protruding endsIf recognized sequence is 5'-GGTACC-3', then generate compatible ends for Anza 34 Pfl23II, Anza 39 Bsp1407I, Anza 91 Acc65IAnza universal protocol
heat inactivate at 80°C for 20 minutes
2-step, heat inactivate first
The target site may have overlapping CpG methylation which will result in blocked or impaired DNA cleavageThe target site may have overlapping Dcm methylation which will result in blocked or impaired DNA cleavage

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