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酶切位点的保护碱基表

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切割率%
2 hr 20 hr
0
0
0
0
0
0
0
0
>90
>90
>90
>90
>90
>90
>90
>90
>90
>90
50
>90
>90
>90
>90
>90
10
25
>90
>90
>90
>90
0
0
75
>90
25
>90
0
0
0
0
50
>90
0
10
0
0
25
50
25
>90
0
0
0
0
>90
>90
50
50
>90
>90
>90
>90
>90
>90
切割率%
2 hr 20 hr
0
0
10
10
10
10
25
90
25
>90
Nsi I
TGCATGCATGCA CCAATGCATTGGTTCTGCAGTT
10
>90
>90
>90
Pac I
TTAATTAA GTTAATTAAC CCTTAATTAAGG
0
0
0
25
0
>90
Pme I
GTTTAAAC GGTTTAAACC GGGTTTAAACCC AGCTTTGTTTAAACGGCGCGCCGG
Hae III Hind III Kpn I
Mlu I Nco I Nde I
Nhe I
GGGGCCCC AGCGGCCGCT TTGCGGCCGCAA
CAAGCTTG CCAAGCTTGG CCCAAGCTTGGG
GGGTACCC GGGGTACCCC CGGGGTACCCCG
GACGCGTC CGACGCGTCG
GAA
Sca I
GAGTACTC AAAAGTACTTTT
10
25
75
75
Sma I
CCCGGG CCCCGGGG CCCCCGGGGG TCCCCCGGGGGA
0
10
0
10
10
50
>90
>90
Spe I
GACTAGTC GGACTAGTCC CGGACTAGTCCG CTAGACTAGTCTAG
10
GGCGCGCC AGGCGCGCCT TTGGCGCGCCAA
CCCCGGGG CCCCCGGGGG TCCCCCGGGGGA
CGGATCCG CGGGATCCCG CGCGGATCCGCG
CAGATCTG GAAGATCTTC GGAAGATCTTCC
GGCGCGCC AGGCGCGCCT TTGGCGCGCCAA
0
0
10
25
0
10
Sac I
CGAGCTCG
10
10
Sac II
GCCGCGGC TCCCCGCGGGGA
0
0
50
>90
Sal I
GTCGACGTCAAAAGGCCATAGCGGCCGC
0
0
GCGTCGACGTCTTGGCCATAGCGGCCGCG
10
50
G
10
75
ACGCGTCGACGTCGGCCATAGCGGCCGCG
CCTCGAGG CCCTCGAGGG CCGCTCGAGCGG
CCCCGGGG CCCCCGGGGG CCCCCCGGGGGG TCCCCCCGGGGGGA
10
50
>90
>90
>90
>90
>90
>90
0
0
>90
>90
75
>90
75
>90
0
0
10
25
10
75
0
0
25
75
50
>90
>90
>90
PCR 设计引物时酶切位点的保护碱基表
酶 Acc I Afl III Asc I Apa I BamH I Bgl II BssH II BstE II BstX I Cla I
EcoR I
寡核苷酸序列
GGTCGACC CGGTCGACCG CCGGTCGACCGG
CACATGTG CCACATGTGG CCCACATGTGGG
>90
10
>90
0
50
0
50
ቤተ መጻሕፍቲ ባይዱ
Sph I
GGCATGCC CATGCATGCATG
0
0
0
25
Stu I Xba I Xho I Xma I
ACATGCATGCATGT
AAGGCCTT GAAGGCCTTC AAAAGGCCTTTT
CTCTAGAG GCTCTAGAGC TGCTCTAGAGCA CTAGTCTAGACTAG
>90
>90
>90
>90
>90
>90
0
0
0
0
10
75
0
0
>90
>90
>90
>90
0
0
25
50
0
0
50
75
0
0
0
0
0
0
0
0
75
>90
75
>90
0
0
10
25
10
50
酶 Not I
寡核苷酸序列
TTGCGGCCGCAA ATTTGCGGCCGCTTTA AAATATGCGGCCGCTATAAA ATAAGAATGCGGCCGCTAAACTAT AAGGAAAAAAGCGGCCGCAAAAGGAAAA
GGGT(A/T)ACCC
AACTGCAGAACCAATGCATTGG AAAACTGCAGCCAATGCATTGGAA CTGCAGAACCAATGCATTGGATGCAT
CATCGATG GATCGATC CCATCGATGG CCCATCGATGGG
GGAATTCC CGGAATTCCG CCGGAATTCCGG
0
0
0
25
0
50
75
>90
Pst I
GCTGCAGC TGCACTGCAGTGCA AACTGCAGAACCAATGCATTGG AAAACTGCAGCCAATGCATTGGAA CTGCAGAACCAATGCATTGGATGCAT
0
0
10
10
>90
>90
>90
>90
0
0
Pvu I
CCGATCGG ATCGATCGAT TCGCGATCGCGA
CCCATGGG CATGCCATGGCATG
CCATATGG CCCATATGGG CGCCATATGGCG GGGTTTCATATGAAACCC GGAATTCCATATGGAATTCC GGGAATTCCATATGGAATTCCC
GGCTAGCC CGGCTAGCCG CTAGCTAGCTAG

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酶切位点所加保护碱基

酶切位点所加保护碱基

100 (2)
99 (2)
LITMUS 29
LITMUS 29
pNEB193
SpeI
SacI
SacI
MluI
2
SacI
BamHI
1
97 (2)
LITMUS 29
HindIII
BglII
3
100 (2)
LITMUS 29
NsiI
BsiWI
2
100 (2)
LITMUS 29
BssHII
BspEI
2
1
100 (1)
8 (2)
LITMUS 39
LITMUS 38
BsrGI
BsrGI
BsrGI
2
1
99 (2)
88 (2)
8
10
12
0
10
10
0
25
50
Not I
TTGCGGCCGCAA
ATTTGCGGCCGCTTTA
AAATATGCGGCCGCTATAAA
ATAAGAATGCGGCCGCTAAACTAT
AAGGAAAAAAGCGGCCGCAAAAGGAAAA
12
16
20
24
28
0
10
10
25
25
0
10
10
90
>90
Nsi I
Enzyme
Base pairs
from End
%Cleavage
Efficiency
Vector
Initial Cut
AatII
3
2
1
88 (2)
100 (2)
95 (2)
NEB保护碱基-各种酶切位点保护碱基

NEB保护碱基-各种酶切位点保护碱基


0
0
0
0
0
0
0
0
75
>90
75
>90
Nhe I
GGCTAGCC CGGCTAGCCG CTAGCTAGCTAG
0
0
10
25
10
50
Not I
TTGCGGCCGCAA
0
0
ATTTGCGGCCGCTTTA
10
10
AAATATGCGGCCGCTATAAA
10
10
ATAAGAATGCGGCCGCTAAACTAT
酶 Acc I Afl III Asc I Ava I BamH I Bgl II BssH II BstE II BstX I Cla I
EcoR I
PCR 设计引物时酶切位点的保护
寡核苷酸序列
GGTCGACC CGGTCGACCG CCGGTCGACCGG
CACATGTG CCACATGTGG CCCACATGTGGG
0
0
0
25
0
50
75
>90
Pst I
GCTGCAGC
0
0
TGCACTGCAGTGCA
10
10
AACTGCAGAACCAATGCATTGG
>90
>90
AAAACTGCAGCCAATGCATTGGAA
>90
>90
CTGCAGAACCAATGCATTGGATGCAT
0
0
Pvu I
CCGATCGG ATCGATCGAT TCGCGATCGCGA
25
90
AAGGAAAAAAGCGGCCGCAAAAGGAAAA
保护碱基列表

保护碱基列表


0
0
75
>90
25
>90
0
0
0
0
50
>90
0
10
0
0
25
50
25
>90
0
0
0
0
>90
>90
50
50
>90
>90
>90
>90
>90
>90
>90
>90
>90
>90
>90
>90
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Hind III
CAAGCTTG CCAAGCTTGG CCCAAGCTTGGG
GGAATTCC CGGAATTCCG CCGGAATTCCGG
GGGGCCCC AGCGGCCGCT TTGCGGCCGCAA
切割率%

2 hr
0 0 0
20 hr
0 0 0
0
0
>90
>90
>90
>90
>90
>90
>90
>90
>90
>90
50
>90
>90
>90
>90
>90
10
25
>90
>90
>90
>90
CACATGTG CCACATGTGG CCCACATGTGGG
GGCGCGCC AGGCGCGCCT TTGGCGCGCCAA
常用酶切位点表(含保护碱基)课件.doc

常用酶切位点表(含保护碱基)课件.doc

切割率%酶寡核苷酸序列2 hr 20 hrAcc I G GTCGAC C 0 0CG GTCGAC CG 0 0CCG GTCGAC CGG 0 0Afl III C ACATGT G 0 0CC ACATGT GG >90 >90CCC ACATGT GGG >90 >90Asc I GGCGCGCC >90 >90A GGCGCGCC T >90 >90TT GGCGCGCC AA >90 >90Ava I C CCCGGG G 50 >90CC CCCGGG GG >90 >90TCC CCCGGG GGA >90 >90BamH I C GGATCC G 10 25CG GGATCC CG >90 >90CGC GGATCC GCG >90 >90Bgl II C AGATCT G 0 0GA AGATCT TC 75 >90GGA AGATCT TCC 25 >90BssH II G GCGCGC C 0 0AG GCGCGC CT 0 0TTG GCGCGC CAA 50 >90BstE II G GGT(A/T)ACC C 0 10BstX I AACTGCAGAA CCAATGCATTGG 0 0 AAAACTGCAG CCAATGCATTGG AA 25 50CTGCAGAA CCAATGCATTGG ATGCAT 25 >90Cla I C ATCGAT G 0 0GATCGAT C 0 0CC ATCGAT GG >90 >90CCC ATCGAT GGG 50 50EcoR I G GAATTC C >90 >90>90 >90CG GAATTC CGCCG GAATTC CGG >90 >90Hae III GG GGCC CC >90 >90AGC GGCC GCT >90 >90TTGC GGCC GCAA >90 >90Hind III C AAGCTT G 0 0CC AAGCTT GG 0 0CCC AAGCTT GGG 10 75Kpn I G GGTACC C 0 0GG GGTACC CC >90 >90CGG GGTACC CCG >90 >90Mlu I G ACGCGT C 0 0CG ACGCGT CG 25 50Nco I C CCATGG G 0 0CATG CCATGG CATG 50 75Nde I C CATATG G 0 0CC CATATG GG 0 0CGC CATATG GCG 0 0GGGTTT CATATG AAACCC 0 0GGAATTC CATATG GAATTCC 75 >90GGGAATTC CATATG GAATTCCC 75 >90Nhe I G GCTAGC C 0 0CG GCTAGC CG 10 25CTA GCTAGC TAG 10 50切割率%酶寡核苷酸序列2 hr 20 hrNot I TT GCGGCCGC AA 0 0ATTT GCGGCCGC TTTA 10 10AAATAT GCGGCCGC TATAAA 10 10 ATAAGAAT GCGGCCGC TAAACTAT 25 90 AAGGAAAAAA GCGGCCGC AAAAGGAAAA 25 >90Nsi I TGC ATGCAT GCA 10 >90 CCA ATGCAT TGGTTCTGCAGTT >90 >90Pac I TTAATTAA 0 0G TTAATTAA C 0 25CC TTAATTAA GG 0 >90Pme I GTTTAAAC 0 0G GTTTAAAC C 0 25GG GTTTAAAC CC 0 50AGCTTT GTTTAAAC GGCGCGCCGG 75 >90Pst I G CTGCAG C 0 0TGCA CTGCAG TGCA 10 10 AA CTGCAG AACCAATGCATTGG >90 >90AAAA CTGCAG CCAATGCATTGGAA >90 >90CTGCAG AACCAATGCATTGGATGCAT 0 0Pvu I C CGATCG G 0 0AT CGATCG AT 10 25TCG CGATCG CGA 0 10 Sac I C GAGCTC G 10 10Sac II GCCGCGG C 0 0TCC CCGCGG GGA 50 >90Sal I GTCGAC GTCAAAAGGCCATAGCGGCCGC 0 0 GC GTCGAC GTCTTGGCCATAGCGGCCGCG 10 50G 10 75ACGC GTCGAC GTCGGCCATAGCGGCCGCGGAASca I GAGTACT C 10 25AAA AGTACT TTT 75 75Sma I CCCGGG 0 10CCCCGGG G 0 10CC CCCGGG GG 10 50TCC CCCGGG GGA >90 >90Spe I GACTAGT C 10 >90GG ACTAGT CC 10 >90CGG ACTAGT CCG 0 50CTAG ACTAGT CTAG 0 50Sph I G GCATGC C 0 0CAT GCATGC ATG 0 25ACAT GCATGC ATGT 10 50Stu I AAGGCCT T >90 >90GA AGGCCT TC >90 >90AAA AGGCCT TTT >90 >90Xba I CTCTAGA G 0 0GC TCTAGA GC >90 >90TGC TCTAGA GCA 75 >90CTAG TCTAGA CTAG 75 >90Xho I C CTCGAG G 0 0CC CTCGAG GG 10 25CCG CTCGAG CGG 10 75Xma I CCCCGGG G 0 0CC CCCGGG GG 25 75CCC CCCGGG GGG 50 >90TCCC CCCGGG GGGA >90 >90。