2024年3月10日发(作者：朱盼晴)
NEB公司提供的保护碱基序列表：
Enzyme
Oligo Sequence
GGTCGACC
CGGTCGACCG
CCGGTCGACCGG
CACATGTG
CCACATGTGG
CCCACATGTGGG
GGCGCGCC
AGGCGCGCCT
TTGGCGCGCCAA
CCCCGGGG
CCCCCGGGGG
TCCCCCGGGGGA
CGGATCCG
CGGGATCCCG
CGCGGATCCGCG
CAGATCTG
GAAGATCTTC
GGAAGATCTTCC
GGCGCGCC
AGGCGCGCCT
TTGGCGCGCCAA
GGGT(A/T)ACCC
AACTGCAGAACCAATGCATTGG
AAAACTGCAGCCAATGCATTGGAA
CTGCAGAACCAATGCATTGGATGCAT
CATCGATG
GATCGATC
CCATCGATGG
CCCATCGATGGG
Oligo Sequence
GGAATTCC
CGGAATTCCG
CCGGAATTCCGG
AccI
AflIII
AscI
AvaI
BamHI
BglII
BssHII
BstEII
BstXI
ClaI
Enzyme
EcoRI
HaeIII
GGGGCCCC
AGCGGCCGCT
TTGCGGCCGCAA
CAAGCTTG
CCAAGCTTGG
CCCAAGCTTGGG
GGGTACCC
GGGGTACCCC
CGGGGTACCCCG
GACGCGTC
CGACGCGTCG
CCCATGGG
CATGCCATGGCATG
CCATATGG
CCCATATGGG
CGCCATATGGCG
GGGTTTCATATGAAACCC
GGAATTCCATATGGAATTCC
GGGAATTCCATATGGAATTCCC
GGCTAGCC
CGGCTAGCCG
CTAGCTAGCTAG
TTGCGGCCGCAA
ATTTGCGGCCGCTTTA
AAATATGCGGCCGCTATAAA
ATAAGAATGCGGCCGCTAAACTAT
AAGGAAAAAAGCGGCCGCAAAAGGAAAA
Oligo Sequence
TGCATGCATGCA
CCAATGCATTGGTTCTGCAGTT
TTAATTAA
GTTAATTAAC
CCTTAATTAAGG
GTTTAAAC
GGTTTAAACC
GGGTTTAAACCC
AGCTTTGTTTAAACGGCGCGCCGG
Chain
Length
12
22
8
10
12
8
10
12
24
HindIII
KpnI
MluI
NcoI
NdeI
NheI
NotI
Enzyme
NsiI
PacI
PmeI
PstI
GCTGCAGC
TGCACTGCAGTGCA
AACTGCAGAACCAATGCATTGG
AAAACTGCAGCCAATGCATTGGAA
CTGCAGAACCAATGCATTGGATGCAT
CCGATCGG
ATCGATCGAT
TCGCGATCGCGA
CGAGCTCG
8
14
22
24
26
8
10
12
8
8
12
28
30
32
8
12
6
8
10
12
PvuI
SacI
SacII
GCCGCGGC
TCCCCGCGGGGA
GTCGACGTCAAAAGGCCATAGCGGCCGC
GCGTCGACGTCTTGGCCATAGCGGCCGCGG
ACGCGTCGACGTCGGCCATAGCGGCCGCGGAA
GAGTACTC
AAAAGTACTTTT
CCCGGG
CCCCGGGG
CCCCCGGGGG
TCCCCCGGGGGA
Chain
Length
8
10
12
14
8
12
14
8
10
12
8
10
12
14
8
% Cleavage
2 hr
10
10
0
0
0
0
10
>90
>90
>90
0
>90
75
75
0
20 hr
>90
>90
50
50
0
25
50
>90
>90
>90
0
>90
>90
>90
0
SalI
ScaI
SmaI
Enzyme
Oligo Sequence
GACTAGTC
GGACTAGTCC
CGGACTAGTCCG
CTAGACTAGTCTAG
GGCATGCC
CATGCATGCATG
ACATGCATGCATGT
AAGGCCTT
GAAGGCCTTC
AAAAGGCCTTTT
CTCTAGAG
GCTCTAGAGC
TGCTCTAGAGCA
CTAGTCTAGACTAG
CCTCGAGG
SpeI
SphI
StuI
XbaI
XhoI
CCCTCGAGGG
CCGCTCGAGCGG
CCCCGGGG
CCCCCGGGGG
CCCCCCGGGGGG
TCCCCCCGGGGGGA
10
12
8
10
12
14
10
10
0
25
50
>90
25
75
0
75
>90
>90
XmaI
四. Fermentas公司提供的关于保护碱基的资料，其实这个更有参考价值~
太多了，请参考：
/en/support/technical-reference/restriction-enzymes/cleavage-e
fficiency
NEB公司的请参考：
/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized
_
/nebecomm/tech_reference/restriction_enzymes/cleavage_olignucleo
#A
Table. Cleavage Efficiency Close to the Termini of PCR Fragments
Enzyme
AanI (PsiI)
AarI
Bp from the recognition site to fragment end
1
0
20-50
50-100
0
2
0-20
0-20
50-100
20-50
20-50
20-50
50-100
3
20-50
50-100
20-50
4
5
AasI (DrdI)
AatII
Acc65I (Asp718I)
AdeI (DraIII)
AjiI (BmgBI)
AluI
0-20
50-100
50-100
0-20
50-100
50-100
0
50-100
50-100
50-100
Alw21I (BsiHKAI)
Alw26I
Alw44I
ApaI
50-100
50-100
50-100
BamHI
BclI
BauI (BssSI)0-20
0
BcnI (NciI)
BcuI (SpeI)
BfiI (BmrI)
BfmI (SfcI)
20-50
50-100
50-100
50-100
50-100
BfuI (BciVI)50-100
BglI20-50
BglII0
Bme1390I (ScrFI)20-50
BoxI (PshAI)0
BpiI (BbsI)50-100
Bpu10I20-50
Bpu1102I (BlpI)50-100
BseDI (BsaJI)0
BseGI (BtsCI)50-100
BseJI (BsaBI)0
BseLI (BslI)0
BseMI (BsrDI)0-20
BseMII (BspCNI)50-100
BseNI (BsrI)0
BseSI (Bme1580I)50-100
BseXI (BbvI)20-50
Bsh1236I (BstUI)50-100
Bsh1285I (BsiEI)0-20
BshNI (BanI)50-100
BshTI (AgeI)20-50
Bsp119I (BstBI)50-100
Bsp120I (PspOMI)20-50
Bsp1407I (BsrGI)20-50
Bsp143I (Sau3AI)50-100
Bsp68I (NruI)0
BspLI (NlaIV)50-100
BspOI (BmtI)0
BspPI (AlwI)0
BspTI (AflII)0
Bst1107I (BstZ17I)0-20
BstXI0
Bsu15I (ClaI)50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
20-50
50-100
0-20
50-100
50-100
50-100
50-100
50-100
BsuRI (HaeIII)
BveI (BspMI)
CaiI (AlwNI)
0-20
0-20
0
20-50
20-50
0-20
50-100
50-100
50-100
50-100
Cfr10I (BsrFI)
Cfr42I (SacII)
Cfr13I (Sau96I)50-100
50-100
Cfr9I (XmaI)20-50
CfrI (EaeI)0
CpoI (RsrII)50-100
CseI (HgaI)50-100
Csp6I (CviQI)50-100
DpnI50-100
DraI0
Eam1104I (EarI)0
Eam1105I (AhdI)0
Ecl136II (EcoICRI)50-100
Eco105I (SnaBI)20-50
Eco130I (StyI)0
Eco147I (StuI)0
Eco24I (BanII)50-100
Eco31I (BsaI)20-50
Eco32I (EcoRV)20-50
Eco47I (AvaII)50-100
Eco47III (AfeI)0
Eco52I (EagI)0-20
Eco57I (AcuI)50-100
Eco57MI50-100
Eco72I (PmlI)0-20
Eco81I (Bsu36I)50-100
Eco88I (AvaI)50-100
Eco91I (BstEII)20-50
EcoO109I (DraII)50-100
EcoRI50-100
EcoRII0
EheI (SfoI)20-50
Esp3I (BsmBI)50-100
FaqI (BsmFI)0-20
50-100
50-100
0-20
50-100
50-100
50-100
50-100
50-100
0-20
50-100
50-100
50-100
0-20
50-100
50-100
50-100
50-100
50-100
20-50
50-100
50-100
FspAI0-20 20-50
50-100
0-20
50-100
FspBI (BfaI)
GsuI (BpmI)
HhaI
20-50
50-100
50-100
Hin1I (BsaHI)50-100
Hin1II (NlaIII)
Hin6I (HinP1I)
0
0
50-100 HincII (HindII)
50-100
0-20 50-100
HindIII
HinfI
HpaII
HphI
Hpy8I (MjaIV)
HpyF10VI (MwoI)
HpyF3I (DdeI)
I-SceI
Kpn2I (BspEI)
KpnI
KspAI (HpaI)
LguI (SapI)
Lsp1109I (BbvI)
LweI (SfaNI)
MauBI
MbiI (BsrBI)
MboI
MboII
MlsI (MscI)
MluI
MnlI
Mph1103I (NsiI)
MreI (Sse232I)
MspI (HpaII)
MssI (PmeI)
MunI (MfeI)
Mva1269I (BsmI)
MvaI (BstNI)
NcoI
0
50-100
0-20
0
0-20
50-100
20-50
50-100
0
50-100
20-50
50-100
0-20
20-50
0
0
50-100
50-100
0-20
20-50
0
20-50
0
0-20
20-50
20-50
0
0
0
0-20
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
0-20
0-20
50-100
50-100
50-100
20-50
50-100
50-100
50-100
50-100
20-50
50-100
50-100
50-100
50-100
50-100
50-100
20-50
NdeI
NheI
NmuCI (Tsp45I)
NotI
NsbI (FspI)
OliI (AleI)
PacI
PaeI (SphI)
PagI (BspHI)
PasI
PauI (BssHII)
PdiI (NaeI)
PdmI (XmnI)
PfeI (TfiI)
Pfl23II (BsiWI)
PfoI
Ppu21I (BsaAI)
PscI (PciI)
Psp1406I (AclI)
Psp5II (PpuMI)
PstI
PsuI (BstYI)
PsyI (Tth111I)
PvuI
PvuII
RsaI
RseI (MslI)
SacI
SalI
SatI (Fnu4HI)
ScaI
SchI (MlyI)
SdaI (SbfI)
SduI (Bsp1286I)
SfaAI (AsiSI)
SfiI
SgrDI
0-20
0
20-50
20-50
0
0-20
20-50
0
20-50
50-100
0
0-20
50-100
50-100
0-20
0
50-100
0
0-20
0
0-20
0-20
0
20-50
50-100
50-100
0
50-100
20-50
0
0-20
50-100
0-20
50-100
0
50-100
0-20
20-50
20-50
50-100
50-100
0-20
20-50
50-100
0-20
50-100
50-100
50-100
20-50
20-50
50-100
50-100
50-100
50-100
50-100
50-100
0-20
50-100
50-100
50-100
0-20
20-50
50-100
50-100
50-100
50-100
50-100
50-100
20-50
50-100
50-100
50-100
20-50
50-100
20-50
50-100
SgsI (AscI)
SmaI
50-100
50-100
0-20
0
50-100
50-100
SmiI (SwaI)
SmoI (SmlI)
SmuI (FauI)
SsiI (AciI)
50-100
50-100
SspDI (KasI)
SspI
TaaI (HpyCH4III)
TaiI (MaeII)
TaqI
TasI (Tsp509I)
TatI
TauI
Tru1I (MseI)
TscAI (TspRI)
TsoI
Van91I (PflMI)
VspI (AseI)
XagI (EcoNI)
XapI (ApoI)
XbaI
XceI (NspI)
XhoI
XmaJI (AvrII)
XmiI (AccI)
20-50
0-20
20-50
50-100
0
0
0-20
20-50
0
0
20-50
0
50-100
20-50
20-50
20-50
0
0-20
50-100
50-100
50-100
50-100
20-50
20-50
50-100
0-20
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
20-50
50-100
50-100
50-100
50-100
2024年3月10日发(作者：朱盼晴)
NEB公司提供的保护碱基序列表：
Enzyme
Oligo Sequence
GGTCGACC
CGGTCGACCG
CCGGTCGACCGG
CACATGTG
CCACATGTGG
CCCACATGTGGG
GGCGCGCC
AGGCGCGCCT
TTGGCGCGCCAA
CCCCGGGG
CCCCCGGGGG
TCCCCCGGGGGA
CGGATCCG
CGGGATCCCG
CGCGGATCCGCG
CAGATCTG
GAAGATCTTC
GGAAGATCTTCC
GGCGCGCC
AGGCGCGCCT
TTGGCGCGCCAA
GGGT(A/T)ACCC
AACTGCAGAACCAATGCATTGG
AAAACTGCAGCCAATGCATTGGAA
CTGCAGAACCAATGCATTGGATGCAT
CATCGATG
GATCGATC
CCATCGATGG
CCCATCGATGGG
Oligo Sequence
GGAATTCC
CGGAATTCCG
CCGGAATTCCGG
AccI
AflIII
AscI
AvaI
BamHI
BglII
BssHII
BstEII
BstXI
ClaI
Enzyme
EcoRI
HaeIII
GGGGCCCC
AGCGGCCGCT
TTGCGGCCGCAA
CAAGCTTG
CCAAGCTTGG
CCCAAGCTTGGG
GGGTACCC
GGGGTACCCC
CGGGGTACCCCG
GACGCGTC
CGACGCGTCG
CCCATGGG
CATGCCATGGCATG
CCATATGG
CCCATATGGG
CGCCATATGGCG
GGGTTTCATATGAAACCC
GGAATTCCATATGGAATTCC
GGGAATTCCATATGGAATTCCC
GGCTAGCC
CGGCTAGCCG
CTAGCTAGCTAG
TTGCGGCCGCAA
ATTTGCGGCCGCTTTA
AAATATGCGGCCGCTATAAA
ATAAGAATGCGGCCGCTAAACTAT
AAGGAAAAAAGCGGCCGCAAAAGGAAAA
Oligo Sequence
TGCATGCATGCA
CCAATGCATTGGTTCTGCAGTT
TTAATTAA
GTTAATTAAC
CCTTAATTAAGG
GTTTAAAC
GGTTTAAACC
GGGTTTAAACCC
AGCTTTGTTTAAACGGCGCGCCGG
Chain
Length
12
22
8
10
12
8
10
12
24
HindIII
KpnI
MluI
NcoI
NdeI
NheI
NotI
Enzyme
NsiI
PacI
PmeI
PstI
GCTGCAGC
TGCACTGCAGTGCA
AACTGCAGAACCAATGCATTGG
AAAACTGCAGCCAATGCATTGGAA
CTGCAGAACCAATGCATTGGATGCAT
CCGATCGG
ATCGATCGAT
TCGCGATCGCGA
CGAGCTCG
8
14
22
24
26
8
10
12
8
8
12
28
30
32
8
12
6
8
10
12
PvuI
SacI
SacII
GCCGCGGC
TCCCCGCGGGGA
GTCGACGTCAAAAGGCCATAGCGGCCGC
GCGTCGACGTCTTGGCCATAGCGGCCGCGG
ACGCGTCGACGTCGGCCATAGCGGCCGCGGAA
GAGTACTC
AAAAGTACTTTT
CCCGGG
CCCCGGGG
CCCCCGGGGG
TCCCCCGGGGGA
Chain
Length
8
10
12
14
8
12
14
8
10
12
8
10
12
14
8
% Cleavage
2 hr
10
10
0
0
0
0
10
>90
>90
>90
0
>90
75
75
0
20 hr
>90
>90
50
50
0
25
50
>90
>90
>90
0
>90
>90
>90
0
SalI
ScaI
SmaI
Enzyme
Oligo Sequence
GACTAGTC
GGACTAGTCC
CGGACTAGTCCG
CTAGACTAGTCTAG
GGCATGCC
CATGCATGCATG
ACATGCATGCATGT
AAGGCCTT
GAAGGCCTTC
AAAAGGCCTTTT
CTCTAGAG
GCTCTAGAGC
TGCTCTAGAGCA
CTAGTCTAGACTAG
CCTCGAGG
SpeI
SphI
StuI
XbaI
XhoI
CCCTCGAGGG
CCGCTCGAGCGG
CCCCGGGG
CCCCCGGGGG
CCCCCCGGGGGG
TCCCCCCGGGGGGA
10
12
8
10
12
14
10
10
0
25
50
>90
25
75
0
75
>90
>90
XmaI
四. Fermentas公司提供的关于保护碱基的资料，其实这个更有参考价值~
太多了，请参考：
/en/support/technical-reference/restriction-enzymes/cleavage-e
fficiency
NEB公司的请参考：
/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized
_
/nebecomm/tech_reference/restriction_enzymes/cleavage_olignucleo
#A
Table. Cleavage Efficiency Close to the Termini of PCR Fragments
Enzyme
AanI (PsiI)
AarI
Bp from the recognition site to fragment end
1
0
20-50
50-100
0
2
0-20
0-20
50-100
20-50
20-50
20-50
50-100
3
20-50
50-100
20-50
4
5
AasI (DrdI)
AatII
Acc65I (Asp718I)
AdeI (DraIII)
AjiI (BmgBI)
AluI
0-20
50-100
50-100
0-20
50-100
50-100
0
50-100
50-100
50-100
Alw21I (BsiHKAI)
Alw26I
Alw44I
ApaI
50-100
50-100
50-100
BamHI
BclI
BauI (BssSI)0-20
0
BcnI (NciI)
BcuI (SpeI)
BfiI (BmrI)
BfmI (SfcI)
20-50
50-100
50-100
50-100
50-100
BfuI (BciVI)50-100
BglI20-50
BglII0
Bme1390I (ScrFI)20-50
BoxI (PshAI)0
BpiI (BbsI)50-100
Bpu10I20-50
Bpu1102I (BlpI)50-100
BseDI (BsaJI)0
BseGI (BtsCI)50-100
BseJI (BsaBI)0
BseLI (BslI)0
BseMI (BsrDI)0-20
BseMII (BspCNI)50-100
BseNI (BsrI)0
BseSI (Bme1580I)50-100
BseXI (BbvI)20-50
Bsh1236I (BstUI)50-100
Bsh1285I (BsiEI)0-20
BshNI (BanI)50-100
BshTI (AgeI)20-50
Bsp119I (BstBI)50-100
Bsp120I (PspOMI)20-50
Bsp1407I (BsrGI)20-50
Bsp143I (Sau3AI)50-100
Bsp68I (NruI)0
BspLI (NlaIV)50-100
BspOI (BmtI)0
BspPI (AlwI)0
BspTI (AflII)0
Bst1107I (BstZ17I)0-20
BstXI0
Bsu15I (ClaI)50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
20-50
50-100
0-20
50-100
50-100
50-100
50-100
50-100
BsuRI (HaeIII)
BveI (BspMI)
CaiI (AlwNI)
0-20
0-20
0
20-50
20-50
0-20
50-100
50-100
50-100
50-100
Cfr10I (BsrFI)
Cfr42I (SacII)
Cfr13I (Sau96I)50-100
50-100
Cfr9I (XmaI)20-50
CfrI (EaeI)0
CpoI (RsrII)50-100
CseI (HgaI)50-100
Csp6I (CviQI)50-100
DpnI50-100
DraI0
Eam1104I (EarI)0
Eam1105I (AhdI)0
Ecl136II (EcoICRI)50-100
Eco105I (SnaBI)20-50
Eco130I (StyI)0
Eco147I (StuI)0
Eco24I (BanII)50-100
Eco31I (BsaI)20-50
Eco32I (EcoRV)20-50
Eco47I (AvaII)50-100
Eco47III (AfeI)0
Eco52I (EagI)0-20
Eco57I (AcuI)50-100
Eco57MI50-100
Eco72I (PmlI)0-20
Eco81I (Bsu36I)50-100
Eco88I (AvaI)50-100
Eco91I (BstEII)20-50
EcoO109I (DraII)50-100
EcoRI50-100
EcoRII0
EheI (SfoI)20-50
Esp3I (BsmBI)50-100
FaqI (BsmFI)0-20
50-100
50-100
0-20
50-100
50-100
50-100
50-100
50-100
0-20
50-100
50-100
50-100
0-20
50-100
50-100
50-100
50-100
50-100
20-50
50-100
50-100
FspAI0-20 20-50
50-100
0-20
50-100
FspBI (BfaI)
GsuI (BpmI)
HhaI
20-50
50-100
50-100
Hin1I (BsaHI)50-100
Hin1II (NlaIII)
Hin6I (HinP1I)
0
0
50-100 HincII (HindII)
50-100
0-20 50-100
HindIII
HinfI
HpaII
HphI
Hpy8I (MjaIV)
HpyF10VI (MwoI)
HpyF3I (DdeI)
I-SceI
Kpn2I (BspEI)
KpnI
KspAI (HpaI)
LguI (SapI)
Lsp1109I (BbvI)
LweI (SfaNI)
MauBI
MbiI (BsrBI)
MboI
MboII
MlsI (MscI)
MluI
MnlI
Mph1103I (NsiI)
MreI (Sse232I)
MspI (HpaII)
MssI (PmeI)
MunI (MfeI)
Mva1269I (BsmI)
MvaI (BstNI)
NcoI
0
50-100
0-20
0
0-20
50-100
20-50
50-100
0
50-100
20-50
50-100
0-20
20-50
0
0
50-100
50-100
0-20
20-50
0
20-50
0
0-20
20-50
20-50
0
0
0
0-20
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
0-20
0-20
50-100
50-100
50-100
20-50
50-100
50-100
50-100
50-100
20-50
50-100
50-100
50-100
50-100
50-100
50-100
20-50
NdeI
NheI
NmuCI (Tsp45I)
NotI
NsbI (FspI)
OliI (AleI)
PacI
PaeI (SphI)
PagI (BspHI)
PasI
PauI (BssHII)
PdiI (NaeI)
PdmI (XmnI)
PfeI (TfiI)
Pfl23II (BsiWI)
PfoI
Ppu21I (BsaAI)
PscI (PciI)
Psp1406I (AclI)
Psp5II (PpuMI)
PstI
PsuI (BstYI)
PsyI (Tth111I)
PvuI
PvuII
RsaI
RseI (MslI)
SacI
SalI
SatI (Fnu4HI)
ScaI
SchI (MlyI)
SdaI (SbfI)
SduI (Bsp1286I)
SfaAI (AsiSI)
SfiI
SgrDI
0-20
0
20-50
20-50
0
0-20
20-50
0
20-50
50-100
0
0-20
50-100
50-100
0-20
0
50-100
0
0-20
0
0-20
0-20
0
20-50
50-100
50-100
0
50-100
20-50
0
0-20
50-100
0-20
50-100
0
50-100
0-20
20-50
20-50
50-100
50-100
0-20
20-50
50-100
0-20
50-100
50-100
50-100
20-50
20-50
50-100
50-100
50-100
50-100
50-100
50-100
0-20
50-100
50-100
50-100
0-20
20-50
50-100
50-100
50-100
50-100
50-100
50-100
20-50
50-100
50-100
50-100
20-50
50-100
20-50
50-100
SgsI (AscI)
SmaI
50-100
50-100
0-20
0
50-100
50-100
SmiI (SwaI)
SmoI (SmlI)
SmuI (FauI)
SsiI (AciI)
50-100
50-100
SspDI (KasI)
SspI
TaaI (HpyCH4III)
TaiI (MaeII)
TaqI
TasI (Tsp509I)
TatI
TauI
Tru1I (MseI)
TscAI (TspRI)
TsoI
Van91I (PflMI)
VspI (AseI)
XagI (EcoNI)
XapI (ApoI)
XbaI
XceI (NspI)
XhoI
XmaJI (AvrII)
XmiI (AccI)
20-50
0-20
20-50
50-100
0
0
0-20
20-50
0
0
20-50
0
50-100
20-50
20-50
20-50
0
0-20
50-100
50-100
50-100
50-100
20-50
20-50
50-100
0-20
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
50-100
20-50
50-100
50-100
50-100
50-100