Thermo Scientific 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 Thermo Scientific Molecular Biology enzymes and used in common applications.

DNA/RNA modifying enzymeFastDigest (Green), Tango 1X and
Tango 2X
BGORBamHIEcl136II, SacIEcoRIKpnITaq with KClTaq with (NH4)2SO4PfuRTT4 DNA Ligase
 Enzyme activity in 1X buffers, %
T4 DNA Ligase*
75-10010010075-
100
75-
100
75-
100
5075-
100
10075757575100
FastAP Thermosensitive Alkaline Phosphatase
1001001001001001001001001001005075-
100
100nd
T4 Polynucleotide Kinase (T4 PNK)**
10075-
100
10010075-
100
10050-7510075-
100
10000100100
DNA Polymerase I
10025-5075-
100
10010010050-7510050-75100100100100nd
Klenow Fragment
10025-5020-5010010010050-7510050-75100100100100100
Klenow Fragment, exo-
10025-5025-5010010010050-7510075-
100
100100100100nd
T4 DNA Polymerase
10075-
100
75-
100
1001001001001001005010050100100
T7 DNA Polymerase
10075-
100
10010010075-
100
75-100100100ndndndndnd
Exonuclease Indndndndndndndndnd100100100ndnd
Exonuclease III0-25 (FastDigest (Green), Tango 2X)

25-50 (Tango 1X)
10025-500-250-250-251000-25100ndndndndnd
Lambda Exonuclease
ndndndndndndndndnd50-7550-7550-
75
ndnd
phi29 DNA Polymerase
10025-10010010010010050-7510050-7525-5075-10075-10075-10075-100
Bsm DNA Polymerase
10025-5075-1001001001000-2510025-5010010075-10010050-75
* 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

B10 mM Tris-HCl
(pH 7.5 at 37°C)
10 mM MgCl2
   0.1 mg/mL BSA
  
G10 mM Tris-HCl
(pH 7.5 at 37°C)
10 mM MgCl250 mM NaCl
  0.1 mg/mL BSA  
O
50 mM Tris-HCl
(pH 7.5 at 37°C)
10 mM MgCl2100 mM NaCl
  0.1 mg/mL BSA  
R
10 mM Tris-HCl
(pH 8.5 at 37°C)
10 mM MgCl2100 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  
BamHI
10 mM Tris-HCl
(pH 8.0 at 37°C)
5 mM MgCl2100 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 MgCl2   0.1 mg/mL BSA  
EcoRI50 mM Tris-HCl
(pH 7.5 at 37°C)
10 mM MgCl2100 mM NaCl
 0.02%
Triton X-100
0.1 mg/mL BSA  
KpnI10 mM Tris-HCl
(pH 7.5 at 37°C)
10 mM MgCl2  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 MgCl250 mM KCl
 0.08%
Nonidet P40
   
Taq with
(NH4)2SO4
75 mM Tris-HCl
(pH 8.8 at 25°C)
2 mM MgCl2  0.01%
Tween 20
 20 mM (NH4)2SO4 
T4 DNA Ligase

140 mM Tris-HCl
(pH 7.8 at 25°C)
10 mM MgCl2 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.

仅供科研使用,不可用于诊断目的。