DNA 聚合酶 I
DNA 聚合酶 I
Invitrogen™

DNA 聚合酶 I

DNA 聚合酶 I 是具有 5´→3´ 和 3´→5´ 脱氧核糖核酸外切酶活性的 DNA 聚合酶了解更多信息
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货号数量
180100251000 U
18010017250 U
货号 18010025
价格（CNY)
6,874.00
Each
添加至购物车
数量:
1000 U
价格（CNY)
6,874.00
Each
添加至购物车
DNA 聚合酶 I 是具有 5´→3´ 和 3´→5´ 脱氧核糖核酸外切酶活性的 DNA 聚合酶。 DNA 聚合酶 I 可掺入生物素化的核苷酸。

应用： 依赖于 DNase I 的切口平移 (1,2)。 cDNA 克隆中的第二链合成 (3)。 填补 5´ 突出端 (4)。

来源： 纯化自表达质粒所带 DNA 聚合酶 I 基因的大肠杆菌。

性能和质量测试： 脱氧核糖核酸内切酶分析；确定依赖于 DNase I 的切口平移的效率。

单位定义： 一个单位的酶量可以在 37°C 的反应体系中，使用模板引物在 30 分钟内向酸沉淀材料掺入 10 nmol 总脱氧核糖核苷酸。

单位反应条件： 反应物共 100 μl，包括 50 mM 磷酸钾 (pH 7.0)、6.7 mM MgCl2、1 mM 2-巯基乙醇、80 μg/ml 模板引物、32 μM dTTP、69 nM [3H]dTTP 和酶，温度为 37°C，时间 30 分钟。
仅供科研使用。不可用于诊断程序。
规格
描述DNA 聚合酶
外切核酸酶活性3' - 5'，5' - 3'
热启动否
聚合酶DNA 聚合酶 I
数量1000 U
运输条件湿冰
Unit SizeEach
内容与储存
1000 单位 DNA 聚合酶，溶于 50 mM 磷酸钾（pH 值 7.0）、100 mM KCl、1 mM DTT 和 50% (v/v) 甘油储存缓冲液中。以 5–10 单位/µL 提供。

在 -5°C 至 -30°C 下储存。

常见问题解答 (FAQ)

Is there anything to prevent AmpliTaq Gold DNA polymerase from extending from the 3’ end of a TaqMan probe in a 5’ nuclease assay?

Yes. There is a phosphate group on the 3' end of all TaqMan probes that prevents such extension.

How does AmpliTaq Gold DNA Polymerase differ from AmpliTaq DNA Polymerase?

AmpliTaq Gold DNA Polymerase is a modified form of AmpliTaq DNA Polymerase that contains a proprietary chemical (or so-called hot start molecule) bound to the enzyme's active site. In order to activate the AmpliTaq Gold DNA Polymerase fully, we recommend an initial activation step of 95 degrees C for 10 min when using GeneAmp 10X PCR Buffer I and/or GeneAmp 10X PCR Buffer II and Mg in one of our thermal cyclers. When using GeneAmp 10X PCR Gold Buffer, activation time can be reduced to 5 minutes. Once activation is complete, you can proceed with your standard PCR cycling program (denaturing, annealing, extension, etc).

Does AmpliTaq Gold DNA Polymerase contain exonuclease (proofreading) activity?

No, AmpliTaq Gold DNA polymerase does not contain proofreading activity, however fidelity in PCR amplifications utilizing this enzyme may be improved. High fidelity can be achieved by: 1. Decreasing the final concentration of each nucleotide to 40-50 uM. 2. Using the lowest MgCl2 concentration possible. 3. Using less enzyme. 4. Decreasing extension times. 5. Using the highest annealing temperature possible. 6. Using as few cycles as possible.

Does the fidelity of AmpliTaq DNA Polymerase change in the presence of base analogs?

The fidelity of this PCR enzyme is affected in two ways. First, AmpliTaq DNA Polymerase typically binds to and incorporates base analogs less efficiently than conventional dNTPs, which means that polymerase activity is lower in reactions that contain base analogs. Second, the analog may pair with more than one conventional complementary template base, so the analog may be incorporated at an increased level compared to conventional dNTPs. For the best fidelity, we recommend that base analogs are included at low concentrations in the reaction.

What is the expected half life of AmpliTaq DNA Polymerase at 95 degrees C?

The half-life of AmpliTaq DNA Polymerase at 95 degrees C is 40 min. During PCR, the sample is only incubated at the programmed temperature for approximately 20 seconds. Therefore, the cycling half-life of AmpliTaq Gold at 95 degrees C is approximately 100 cycles.

Example: AmpliTaq DNA Polymerase experiences about 20 seconds at 95 degrees C per PCR cycle. The t1/2 is at least 33 minutes; (35-40 min). Therefore, 33 min/20 sec/cycle = 100 cycles. 100 PCR cycles reduces enzyme activity by 50%.

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