Therminator II DNA Polymerase

• 产品编号：M0266L      品牌：New England Biolabs       原厂货号：M0266L
• 产品分类：克隆 > 聚合酶 > DNA聚合酶
• 应用分类：

描述：

• Enhanced incorporation of modified nucleotides, especially ribonucleotides

• Isolated from recombinant source

• Supplied with 10X Reaction Buffer

Description: 
Therminator™ II DNA Polymerase is a 9°N™ DNA Polymerase variant with an enhanced ability to incorporate modified substrates such as dideoxynucleotides, ribonucleotides and acyclonucleotides (1,2). 

Therminator II DNA Polymerase was derived from Therminator DNA Polymerase and differs in having one additional amino acid change. This change allows more efficient incorporation of ribonucleotides (1) and nucleotides with modified 3´ functional groups (3).

Source:
An E. coli strain that carries the 9°N (D141A / E143A / A485L / Y409V) DNA Polymerase gene, a genetically engineered form of the native DNA polymerase fromThermococcus species 9°N-7.

Applications:

• DNA sequencing by partial ribosubstitution (4)
• DNA sequencing using dideoxy (5) or acyclo (6) chain terminators
• SNP analysis with dideoxy or acyclo chain terminators (7)

Reagents Supplied: 
ThermoPol® Reaction Buffer Pack (10X) 


3´ to 5´ Exonuclease: No
5´ to 3´ Exonuclease: No
Strand Displacement: Yes

Heat Inactivation:
No


Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in a total reaction volume in 30 minutes at 75°C.

Unit Assay Conditions: 1X ThermoPol Reaction Buffer, 200 µM dNTPs including [³H]-dTTP and 15 nM primed single-stranded M13mp18.

Reaction Conditions: 1X ThermoPol Reaction Buffer, DNA template, primer, 200 µM dNTPs and 0.5–2 units of Therminator II DNA polymerase in a total reaction volume of 100 µl.

1X ThermoPol Reaction Buffer:
20 mM Tris-HCl
10 mM (NH₄)₂SO₄
10 mM KCl
2 mM MgSO₄
0.1% Triton X-100
pH 8.8 @ 25°C

Concentration:
2,000 units/ml

Storage Conditions:
10 mM Tris-HCl
100 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C
 

原厂资料：

• Enhanced incorporation of modified nucleotides, especially ribonucleotides

• Isolated from recombinant source

• Supplied with 10X Reaction Buffer

Description: 
Therminator™ II DNA Polymerase is a 9°N™ DNA Polymerase variant with an enhanced ability to incorporate modified substrates such as dideoxynucleotides, ribonucleotides and acyclonucleotides (1,2). 

Therminator II DNA Polymerase was derived from Therminator DNA Polymerase and differs in having one additional amino acid change. This change allows more efficient incorporation of ribonucleotides (1) and nucleotides with modified 3´ functional groups (3).

Source:
An E. coli strain that carries the 9°N (D141A / E143A / A485L / Y409V) DNA Polymerase gene, a genetically engineered form of the native DNA polymerase fromThermococcus species 9°N-7.

Applications:

• DNA sequencing by partial ribosubstitution (4)
• DNA sequencing using dideoxy (5) or acyclo (6) chain terminators
• SNP analysis with dideoxy or acyclo chain terminators (7)

Reagents Supplied: 
ThermoPol® Reaction Buffer Pack (10X) 


3´ to 5´ Exonuclease: No
5´ to 3´ Exonuclease: No
Strand Displacement: Yes

Heat Inactivation:
No


Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in a total reaction volume in 30 minutes at 75°C.

Unit Assay Conditions: 1X ThermoPol Reaction Buffer, 200 µM dNTPs including [³H]-dTTP and 15 nM primed single-stranded M13mp18.

Reaction Conditions: 1X ThermoPol Reaction Buffer, DNA template, primer, 200 µM dNTPs and 0.5–2 units of Therminator II DNA polymerase in a total reaction volume of 100 µl.

1X ThermoPol Reaction Buffer:
20 mM Tris-HCl
10 mM (NH₄)₂SO₄
10 mM KCl
2 mM MgSO₄
0.1% Triton X-100
pH 8.8 @ 25°C

Concentration:
2,000 units/ml

Storage Conditions:
10 mM Tris-HCl
100 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C