酶切位点保护碱基-PCR引物设计用于限制性内切酶

酶

寡核苷酸序列

切割率 %

2 hr

20 hr

Acc I

G GTCGAC C
CG GTCGAC CG
CCG GTCGAC CGG

0
0
0

0
0
0

Afl III

C ACATGT G
CC ACATGT GG
CCC ACATGT GGG

0
>90
>90

0
>90
>90

Asc I

GGCGCGCC
A GGCGCGCC T
TT GGCGCGCC AA

>90
>90
>90

>90
>90
>90

Ava I

C CCCGGG G
CC CCCGGG GG
TCC CCCGGG GGA

50
>90
>90

>90
>90
>90

BamH I

C GGATCC G
CG GGATCC CG
CGC GGATCC GCG

10
>90
>90

25
>90
>90

Bgl II

C AGATCT G
GA AGATCT TC
GGA AGATCT TCC

0
75
25

0
>90
>90

BssH II

G GCGCGC C
AG GCGCGC CT
TTG GCGCGC CAA

0
0
50

0
0
>90

BstE II

G GGT(A/T)ACC C

0

10

BstX I

AACTGCAGAA CCAATGCATTGG
AAAACTGCAG CCAATGCATTGG AA
CTGCAGAA CCAATGCATTGG ATGCAT

0
25
25

0
50
>90

Cla I

C ATCGAT G
G ATCGAT C
CC ATCGAT GG
CCC ATCGAT GGG

0
0
>90
50

0
0
>90
50

EcoR I

G GAATTC C
CG GAATTC CG
CCG GAATTC CGG

>90
>90
>90

>90
>90
>90

Hae III

GG GGCC CC
AGC GGCC GCT
TTGC GGCC GCAA

>90
>90
>90

>90
>90
>90

Hind III

C AAGCTT G
CC AAGCTT GG
CCC AAGCTT GGG

0
0
10

0
0
75

Kpn I

G GGTACC C
GG GGTACC CC
CGG GGTACC CCG

0
>90
>90

0
>90
>90

Mlu I

G ACGCGT C
CG ACGCGT CG

0
25

0
50

Nco I

C CCATGG G
CATG CCATGG CATG

0
50

0
75

Nde I

C CATATG G
CC CATATG GG
CGC CATATG GCG
GGGTTT CATATG AAACCC
GGAATTC CATATG GAATTCC
GGGAATTC CATATG GAATTCCC

0
0
0
0
75
75

0
0
0
0
>90
>90

Nhe I

G GCTAGC C
CG GCTAGC CG
CTA GCTAGC TAG

0
10
10

0
25
50

Not I

TT GCGGCCGC AA
ATTT GCGGCCGC TTTA
AAATAT GCGGCCGC TATAAA
ATAAGAAT GCGGCCGC TAAACTAT
AAGGAAAAAA GCGGCCGC AAAAGGAAAA

0
10
10
25
25

0
10
10
90
>90

Nsi I

TGC ATGCAT GCA
CCA ATGCAT TGGTTCTGCAGTT

10
>90

>90
>90

Pac I

TTAATTAA
G TTAATTAA C
CC TTAATTAA GG

0
0
0

0
25
>90

Pme I

GTTTAAAC
G GTTTAAAC C
GG GTTTAAAC CC
AGCTTT GTTTAAAC GGCGCGCCGG

0
0
0
75

0
25
50
>90

Pst I

G CTGCAG C
TGCA CTGCAG TGCA
AA CTGCAG AACCAATGCATTGG
AAAA CTGCAG CCAATGCATTGGAA
CTGCAG AACCAATGCATTGGATGCAT

0
10
>90
>90
0

0
10
>90
>90
0

Pvu I

C CGATCG G
AT CGATCG AT
TCG CGATCG CGA

0
10
0

0
25
10

Sac I

C GAGCTC G

10

10

Sac II

G CCGCGG C
TCC CCGCGG GGA

0
50

0
>90

Sal I

GTCGAC GTCAAAAGGCCATAGCGGCCGC
GC GTCGAC GTCTTGGCCATAGCGGCCGCGG
ACGC GTCGAC GTCGGCCATAGCGGCCGCGGAA

0
10
10

0
50
75

Sca I

G AGTACT C
AAA AGTACT TTT

10
75

25
75

Sma I

CCCGGG
C CCCGGG G
CC CCCGGG GG
TCC CCCGGG GGA

0
0
10
>90

10
10
50
>90

Spe I

G ACTAGT C
GG ACTAGT CC
CGG ACTAGT CCG
CTAG ACTAGT CTAG

10
10
0
0

>90
>90
50
50

Sph I

G GCATGC C
CAT GCATGC ATG
ACAT GCATGC ATGT

0
0
10

0
25
50

Stu I

A AGGCCT T
GA AGGCCT TC
AAA AGGCCT TTT

>90
>90
>90

>90
>90
>90

Xba I

C TCTAGA G
GC TCTAGA GC
TGC TCTAGA GCA
CTAG TCTAGA CTAG

0
>90
75
75

0
>90
>90
>90

Xho I

C CTCGAG G
CC CTCGAG GG
CCG CTCGAG CGG

0
10
10

0
25
75

Xma I

C CCCGGG G
CC CCCGGG GG
CCC CCCGGG GGG
TCCC CCCGGG GGGA

0
25
50
>90

0
75
>90
>90