Restriction Enzyme Activity in Each Reaction Buffer

 Relative Activity (%)
Restriction Enzyme
Cut Site
Buffer L
Buffer M
Buffer H
Buffer K
Buffer T (+BSA)
Buffer
AccI
5'-GT↓(Py)(Pu)AC-3'
3'-CA(Pu)(Py)↑TG-5'		20
100
<20
<20*
160
-
AfaI (RsaI)
5'-GT↓AC-3'
3'-CA↑TG-5'		60
60
40
60
100
-
AIuI
5'-AG↓CT-3'
3'-TC↑AG-5'		100
100
<20
40
200
-
ApaI
5'-GGGCC↓C-3'
3'-C↑CCGGG-5'		100
<20
<20
<20
<20
-
BaII
5'-TGG↓CCA-3'
3'-ACC↑GGT-5'		20*
20*
<20*
<20*
40*
100 (Ball buffer)
BamHI
5'-G↓GATCC-3'
3'-CCTAG↑G-5'		<20*
<20
40
100
<20*
-
BgIII
5'-A↓GATCT-3'
3'-TCTAG↑A-5'		<20
20
100
100*
60*
-
BmeT120I (AvaI)
5'-C↓(Py)CG(Pu)G-3'
3'-G(Pu)GC(Py)↑C-5'		<20
<20
20
100
<20
-
BshTI (AgeI)
5'-A↓AGCTT-3'
3'-TTCGA↑A-5'		<20
20
80*
50
50
-
CIaI
5'-AT↓CGAT-3'
3'-TAGC↑TA-5'		40
100
120
100
60
-
DdeI
5'-C↓TNAG-3'
3'-GANT↑C-5'		60
80
100
100
80
-
DpnI
5'-GmA↓TC-3'
3'-CT↑mAG-5'		60
60
120
140
100*
-
DraI (AhaIII)
5'-TTT↓AAA-3'
3'-AAA↑TTT-5'		100
100
60
100
80
-
EcoRI
5'-G↓AATTC-3'
3'-CTTAA↑G-5'		20*
100*
100
120*
80*
-
EcoRV
5'-GAT↓ATC-3'
3'-CTA↑TAG-5'		<20*
40*
100
120*
40*
-
EcoT22I (Ava II)
5'-ATGCA↓T-3'
3'-T↑ACGTA-5'		<20
20
100
140*
20*
-
HaeIII
5'-GG↓CC-3'
3'-CC↑GG-5'		60
100
100
60
100
-
HapII (HpaII)
5'-C↓CGG-3'
3'-GGC↑G-5'		100
60
<20
<20
100
-
HhaI
5'-GCG↓C-3'
3'-C↑GCG-5'		80
100
100
120
120
-
HincII (HindII)
5'-GT(Py)↓(Pu)AC-3'
3'-CA(Pu)↑(Py)TG-5'		20
100
20
40
100
-
HindIII
5'-A↓AGCTT-3'
3'-TTCGA↑A-5'		60*
100
<20
200
100*
-
HinfI
5'-G↓ANTC-3'
3'-CT-↑G-5'		80
100
100
160
60
-
HpaI
5'-GTT↓AAC-3'
3'-CAA↑TTG-5'		<20
40*
20
100
80*
-
KpnI
5'-GGTAC↓C-3'
3'-C↑CATGG-5'		100
60
<20
<20
100*
-
MboI (Sau3AI)
5'-↓GATC-3'
3'-CTAG↑-5'		20
40
60
100
40
-
MIuI
5'-A↓CGCGT-3'
3'-TGCGC↑A-5'		60
60
100
100*
60
-
MseI
5'-T↓TAA-3'
3'-AAT↑T-5'		40
40
20
40
100
-
NcoI
5'-C↓CATGG-3'
3'-GGTAC↑C-5'		40*
60*
20
60*
60*
-
NdeI
5'-CA↓TATG-3'
3'-GTAT↑AC-5'		<20
40
100
10
80
-
NheI
5'-G↓CTAGC-3'
3'-CGATC↑G-5'		120*
100
<20
<20
160*
-
NotI
5'-GC↓GGCCGC-3'
3'-CGCCGG↑CG-5'		<20*
<20*
20**
<20
<20*
-
NruI
5'-TCG↓CGA-3'
3'-AGC↑GCT-5'		0*
<20*
20*
20*
<20*
100 (NrUI buffer)
PstI
5'-CTGCA↓G-3'
3'-G↑ACGTC-5'		<20*
60*
100
80
20*
-
PvuI
5'-CGAT↓CG-3'
3'-GC↑TAGC-5'		<20*
20*
40*
80**
40*
-
PvuII
5'-CAG↓CTG-3'
3'-GTC↑GAC-5'		80*
100
40
<20
40*
-
SacI
5'-GAGCT↓C-3'
3'-C↑TCGAG-5'		100
60
<20
<20
80
-
SacII
5'-CCGC↓GG-3'
3'-GG↑CGCC-5'		40
20
<20
<20
100
-
SaII
5'-G↓TCGAC-3'
3'-CAGCT↑G-5'		<20
<20
100
20*
<20
-
ScaI
5'-AGT↓ACT-3'
3'-TCA↑TGA-5'		<20*
<20*
100
60*
<20*
-
SmaI
5'-CCC↓GGG-3'
3'-GGG↑CCC-5'		<20
<20
<20
<20
100
-
SpeI
5'-A↓CTAGT-3'
3'-TGATC↑A-5'		80*
100
80
100
80*
-
SphI
5'-GCATG↓C-3'
3'-C↑GTACG-5'		20*
40*
100
120
20*
-
SspI
5'-AAT↓ATT-3'
3'-TTA↑TAA-5'		<20*
60*
40
100*
80*
100 (SSpl buffer)
StuI
5'-AGC↓CCT-3'
3'-TCC↑GGA-5'		60
100
60
80
140
-
TaqI (TthHB8I)
5'-T↓CGA-3'
3'-AGC↑T-5'		40*
80*
60*
60*
80*
100 (TaqI buffer)
XbaI
5'-T↓CTAGA-3'
3'-AGATC↑T-5'		<20
80*
20
<20
120
-
XhoI (PaeR7I)
5'-C↓TCGAG-3'
3'-GAGCT↑C-5'		<20
60
100
160
60
-

*Weak star activity is detected
**100% activity is obtained by addition of 0.01% BSA

Buffer Compositions

10X Buffer H
500 mM Tris-HCl, pH 7.5
100 mM MgCl2
10 mM dithiothreitol (DTT)
1000 mM NaCl

10X Buffer K
200 mM Tris-HCl, pH 7.5
100 mM MgCl2
10 mM dithiothreitol (DTT)
1000 mM KCl		10X Buffer L
100 mM Tris-HCl, pH 7.5
100 mM MgCl2
10 mM dithiothreitol (DTT)

10X Buffer M
100 mM Tris-HCl, pH 7.5
100 mM MgCl2
10 mM dithiothreitol (DTT)
500 mM NaCl
10X Buffer T (BSA-free)
330 mM Tris-acetate, pH 7.9
100 mM magnesium acetate
5 mM dithiothreitol (DTT)
660 mM potassium acetate