产品组分信息

本产品提供以下试剂或组分：

NEB #	名称	组分货号	储存温度	数量	浓度
M0274S     -20       T7 DNA Polymerase (unmodified) M0274SVIAL -20 1 x 0.03 ml 10,000 units/ml   T7 DNA Polymerase Reaction Buffer B0274AVIAL -20 1 x 1.5 ml 10 X   Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml
M0274L     -20       T7 DNA Polymerase (unmodified) M0274LVIAL -20 1 x 0.15 ml 10,000 units/ml   T7 DNA Polymerase Reaction Buffer B0274AVIAL -20 1 x 1.5 ml 10 X   Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml

单位定义

One unit is defined as the amount of enzyme that will incorporate10 nmoles of dNTP into acid insoluble material in 30 minutes at 37°C .

反应条件

1X T7 DNA Polymerase Reaction Buffer
Supplement with Recombinant Albumin, Molecular Biology Grade
Incubate at 37°C

1X T7 DNA Polymerase Reaction Buffer
20 mM Tris-HCl
10 mM MgCl₂
1 mM DTT
(pH 7.5 @ 25°C)

贮存溶液

50 mM KPO₄
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7 @ 25°C

热失活

75°C for 20 min

5′ – 3′ 核酸外切酶

No

3′ – 5′ 核酸外切酶

Yes

链置换

No

单位活性检测条件

100 mM KCl, 20 mM Tris-HCl (pH 7.6), 6 mM MgCl₂ , 0.5 mM DTT, 0.1 mM EDTA, 50 ug/ml Recombinant Albumin, 150 μM dNTPs including [³H]-dTTP and 70 µg/ml denatured herring sperm DNA.

错配率

~ 15×10^-6bases

应用特性

• Second strand synthesis in site-directed mutagenesis protocols (8).

相关产品

• Deoxynucleotide (dNTP) Solution Mix
• Deoxynucleotide (dNTP) Solution Set
• Monarch® Plasmid Miniprep Kit 
• Monarch® DNA Gel Extraction Kit
• Monarch® PCR & DNA Cleanup Kit (5 μg)

单独销售的组分

• 重组白蛋白, 分子生物学级（不含动物成分）

1. The high polymerization rate of the enzyme makes long incubations unnecessary.
2. T7 DNA Polymerase is not suitable for DNA sequencing.

1. Hori, K. et al. (1979). J. Biol. Chem.. 254, 11598-11604.
2. Engler, M.J. et al. (1983). J. Biol. Chem.. 258, 11165-11173.
3. Nordstrom, B. et al. (1981). J. Biol. Chem.. 256, 3112-3117.
4. Studier, F.W. et al. (1990). Goeddel, D.V.(Ed.), In Methods Enzymol.. 185, 60-89. San Diego: Academic Press.
5. Grippo, P. and Richardson, C.C. (1971). J. Biol. Chem. 246, 6867-6873.
6. Modrich, P. and Richardson, C.C. (1975). J. Biol. Chem. 250, 5515-5522.
7. Adler, S. and Modrich, P. (1979). J. Biol. Chem. 254, 11605-11614.
8. Bebenek, K. and Kunkel, T.A. (1989). Nucl. Acids Res.. 17, 5408.
9. Mattila, P., et al. (1991). Nucleic Acids Res.. 19, 4967-4973.

• Activity of DNA Modifying Enzymes in rCutSmart™ Buffer

• DNA Polymerase Selection Chart

1. Can T7 DNA Polymerase be used in other NEBuffers, including rCutSmart?
2. Can T7 DNA Polymerase be used to blunt DNA?
3. Can T7 DNA Polymerase be used to fill in 3′ overhangs?
4. Can T7 DNA Polymerase be used to remove 5′ overhangs?
5. Can T7 DNA Polymerase be heat inactivated?
6. Are deoxynucleotides needed to remove a 3′ overhang using T7 DNA Polymerase?
7. When should native T7 DNA Polymerase be used?
8. What is the oligonucleotide-directed mutagenesis without phenotypic selection method?
9. Can the native T7 DNA Polymerase be used for sequencing?
10. Does NEB carry modified T7 Polymerase?
11. Can T7 DNA Polymerase be used in labeling reactions and partial fill in reactions?
12. Are NEB DNA Polymerases supplied with dNTPs?

每一新批次的 NEB 产品都要经过质控，以满足指定的质量标准，对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息，可从此处查阅。

产品说明与变更通知

产品说明表包含产品的储存温度、有效期和规格，这些文件的命名规则如下：[货号]_[规格]_[版本]

• M0274S_L_v1

CoA 文件包含单个批次产品的储存温度、有效期和质控，这些文件的命名规则如下： [货号]_[规格]_[版本]_[Lot#]

• M0274S_L_v1_0111508
• M0274S_L_v1_0111603
• M0274S_L_v1_0111609
• M0274S_L_v1_0111703
• M0274S_L_v1_0111803
• M0274L_v1_10012703
• M0274S_v1_10021765
• M0274L_v1_10021767
• M0274L_v1_10039918
• M0274S_v1_10040917
• M0274L_v1_10057374
• M0274S_v1_10059218
• M0274L_v1_10075649
• M0274L_v1_10081788
• M0274S_v1_10091672
• M0274L_v1_10091094
• M0274S_v1_10107595
• M0274S_v1_10114913
• M0274S_v1_10123097
• M0274L_v1_10131164
• M0274S_v1_10135910
• M0274L_v1_10139934
• M0274L_v1_10152153
• M0274S_v1_10161568
• M0274L_v1_10161623
• M0274S_v1_10163746

以下 SDS 文件可以帮助您安全地使用该产品

• T7 DNA Polymerase (unmodified)
• T7 DNA Polymerase Reaction Buffer
• 重组白蛋白, 分子生物学级