Modifying Enzyme Buffers | Thermo Fisher Scientific

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 enzyme	FastDigest (Green), Tango 1X and Tango 2X	B	G	O	R	BamHI	Ecl136II, SacI	EcoRI	KpnI	Taq with KCl	Taq with (NH₄)₂SO₄	Pfu	RT	T4 DNA Ligase
	Enzyme activity in 1X buffers, %
T4 DNA Ligase*	75-100	100	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	50	75- 100	100	nd
T4 Polynucleotide Kinase (T4 PNK)**	100	75- 100	100	100	75- 100	100	50-75	100	75- 100	100	0	0	100	100
DNA Polymerase I	100	25-50	75- 100	100	100	100	50-75	100	50-75	100	100	100	100	nd
Klenow Fragment	100	25-50	20-50	100	100	100	50-75	100	50-75	100	100	100	100	100
Klenow Fragment, exo-	100	25-50	25-50	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	50	100	50	100	100
T7 DNA Polymerase	100	75- 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	100	100	100	nd	nd
Exonuclease III	0-25 (FastDigest (Green), Tango 2X) 25-50 (Tango 1X)	100	25-50	0-25	0-25	0-25	100	0-25	100	nd	nd	nd	nd	nd
Lambda Exonuclease	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	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	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 MgCl₂				0.1 mg/mL BSA
G	10 mM Tris-HCl (pH 7.5 at 37°C)	10 mM MgCl₂	50 mM NaCl			0.1 mg/mL BSA
O	50 mM Tris-HCl (pH 7.5 at 37°C)	10 mM MgCl₂	100 mM NaCl			0.1 mg/mL BSA
R	10 mM Tris-HCl (pH 8.5 at 37°C)	10 mM MgCl₂	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
BamHI	10 mM Tris-HCl (pH 8.0 at 37°C)	5 mM MgCl₂	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 MgCl₂				0.1 mg/mL BSA
EcoRI	50 mM Tris-HCl (pH 7.5 at 37°C)	10 mM MgCl₂	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 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 MgCl₂	50 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.