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Buffer Compatibility for Modifying Enzymes to print - to print

Fermentas DNA/RNA modifying enzymes are supplied with optimized reaction buffers.
However, it is often convenient to use these enzymes in other buffers for experiments that involve multiple enzymatic reactions.
The table below lists activities of DNA/RNA modifying enzymes in common reaction buffers, supplied with Fermentas enzymes and used in common applications.
 
DNA/RNA modifying enzyme  Enzyme activity in 1X buffers, %
FastDigest®
B
G
O
R
Tango
1X
Tango
2X
BamHI
Ecl136II,
SacI
EcoRI
KpnI
Taq
with KCl
Taq with
(NH4)2SO4
Pfu
 RT T4 DNA
Ligase
T4 DNA Ligase *
75-100 100 100 75-
100
75-
100
75-
100
75-
100
75-
100
50 75-
100
100 75 75 75 75 100
FastAP™ Thermosensitive Alkaline Phosphatase
100 100 100 100 100 100 100 100 100 100 100 100 50 75-
100
100 nd
Shrimp Alkaline Phosphatase
100 100 100 100 100 100 100 100 75-100 100 100 100 100 100 100 nd
T4 Polynucleotide Kinase **
100 75-
100
100 100 75-
100
100 100 100 50-75 100 75-
100
100 0 0 100 100
DNA Polymerase I
100 25-50 75-
100
100 100 100 100 100 50-75 100 50-75 100 100 100 100 nd
Klenow Fragment
100 25-50 20-50 100 100 100 100 100 50-75 100 50-75 100 100 100 100 100
Klenow Fragment, exo-
100 25-50 25-50 100 100 100 100 100 50-75 100 75-
100
100 100 100 100 nd
T4 DNA Polymerase
100 75-
100
75-
100
100 100 100 100 100 100 100 100 50 100 50 100 100
T7 DNA Polymerase
100 75-
100
100 100 100 100 100 75-
100
75-100 100 100 nd nd nd nd nd
Exonuclease I nd nd nd nd nd nd nd nd nd nd nd 100 100 100 nd nd
Exonuclease III 0-25 100 25-50 0-25 0-25 25-50 0-25 0-25 100 0-25 100 nd nd nd nd nd
Lambda Exonuclease
nd nd nd nd nd nd nd nd nd nd nd 50-75 50-75 50-
75
nd nd
phi29 DNA Polymerase
 100 25-100
100
 100 100
100
 100  100 50-75
100
50-75
25-50
 75-100 75-100
 75-100 75-100
Bsm DNA Polymerase
 100 25-50
 75-100 100
 100  100  100 100
 0-25  100  25-50  100  100  75-100  100 50-75
* – buffers were supplemented with 0.5 mM ATP, which is required for T4 DNA Ligase activity.
** – the activity of this enzyme was compared to its activity in buffer A (for forward reaction).
nd – not determined.
 

1X Buffer Composition

B 10 mM Tris-HCl
(pH 7.5 at 37°C) 
10 mM Mgl2
      0.1 mg/ml BSA
   
G 10 mM Tris-HCl
(pH 7.5 at 37°C) 
10 mM Mgl2 50 mM NaCl
    0.1 mg/ml BSA    
O
50 mM Tris-HCl
(pH 7.5 at 37°C)
10 mM Mgl2 100 mM NaCl
    0.1 mg/ml BSA    
R
10 mM Tris-HCl
(pH 8.5 at 37°C)
10 mM Mgl2 100 mM KCl
    0.1 mg/ml BSA    
Tango™
33 mM Tris-acetate
(pH 7.9 at 37°C)
10 mM Mg-acetate
66 mM K-acetate
    0.1 mg/ml BSA    
2X Tango™
66 mM Tris-acetate
(pH 7.9 at 37°C)
20 mM Mg-acetate
132 mM K-acetate
    0.2 mg/ml BSA    
BamHI
10 mM Tris-HCl
(pH 8.0 at 37°C)
5 mM Mgl2 100 mM KCl
  0.02%
Triton X-100
0.1 mg/ml BSA   1 mM BME
Ecl136II,
SacI
10 mM Bis-Tris
Propane-HCl
(pH 6.5 at 37°C)
10 mM Mgl2       0.1 mg/ml BSA    
EcoRI
50 mM Tris-HCl
(pH 7.5 at 37°C)
10 mM Mgl2 100 mM NaCl
  0.02%
Triton X-100
0.1 mg/ml BSA    
KpnI
10 mM Tris-HCl
(pH 7.5 at 37°C)
10 mM Mgl2     0.02%
Triton X-100
0.1 mg/ml BSA    
Taq
with KCl
10 mM Tris-HCl
(pH 8.8 at 25°C)
1.5 mM Mgl2 50 mM KCl
  0.08%
Nonidet P40
     
Taq with
(NH4)2SO4
75 mM Tris-HCl
(pH 8.8 at 25°C)
2 mM Mgl2     0.01%
Tween 20
  20 mM (NH4)2SO4  
Pfu
20 mM Tris-HCl
(pH 8.8 at 25°C)
2 mM MgSO4 10 mM KCl
  0.1%
Triton X-100
0.1 mg/ml BSA 10 mM (NH4)2SO4
 
RT
50 mM Tris-HCl
(pH 8.3 at 25°C)
4 mM Mgl2 50 mM KCl
        10 mM DTT
T4 DNA
Ligase
140 mM Tris-HCl
(pH 7.8 at 25°C)
10 mM Mgl2   0.5 mM ATP
      10 mM DTT

DNA/RNA Modifying Enzyme Dilution

DNA/RNA modifying enzymes are supplied in their optimal storage buffers which are specially formulated for long term storage. If required for specific applications, dilute the enzyme with 1X reaction buffer for short term use.
 
 
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tel: 905 333 8355
fax: 905 333 3135
fermentas.info@thermofisher.com

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