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产品说明:
Hotstart Taq DNA Polymerase High-concentration是一种适体基团修饰的耐热性TAQ DNA聚合酶,高温加热前,适体修饰基团与Taq酶结合,抑制聚合酶的活性,避免了引物延伸的非特异性扩增或引物二聚体的产生,增强了DNA扩增的特异性、灵敏度和稳定性,可广泛适用于常规PCR、多重PCR及巢式PCR等。经过95℃热激10min后,该酶即可恢复活性。此外 Hotstart Taq DNA Polymerase High-concentration没有检测到3'→5'校对外切酶活性,但具有5'→3'外切酶活性,可用于荧光定量PCR的检测。Hotstart Taq DNA Polymerase High-concentration在常温下没有活性,便于PCR实验的常温操作。
产品内容:
1. Hotstart Taq DNA Polymerase High-concentration (10 U/μl )
2. 5×Hotstart Taq Buffer( Mg²⁺ Plus)
3. Solution I (10×)
活性定义:74℃,30 min,使 10 nmol dNTP 掺入酸不溶性沉淀物所需要的酶量定义为 1 个活性单位。
使用方法:
1. PCR 反应体系的设置:
a. 溶解并混匀 PCR 反应所需的各种溶液。并放置于冰浴上或冰盒内。建议反应 PCR 液体分装使用,避免反复冻融。
b. 参考下表设置 PCR 反应,建议 PCR 反应体系的配置在冰浴或在冰盒上进行:

※ 模板 DNA 用量:为了保证反应的灵敏性,25 μl 体系使用 10⁴ 拷贝的靶序列作为模板,可参考下表计算需加入 PCR 体系的模板量。
1 μg 各种来源的 DNA 对应的摩尔数

例如:纯化的人类基因组 DNA 浓度为 1 μg/μl,某基因在人类基因组中拷贝数为 10,其单位体积拷贝数为:
3.0× 10⁵ mol/μg × 1 μg/μl × 10 copy/mol
=3.0× 10⁶ copy/μl
1× 10⁴ copy/ (3.0× 10⁶ copy/μl)
= 1/300 μl
即:1/300 ul 浓度为 1 ug/μl 的人类基因组 DNA 中含 10⁴ 拷贝该基因,稀释 300 倍后加 1μl 至 25 μl PCR 体系。为保证反应的特异性,DNA 终浓度应小于 10 ng/ul,过多的 DNA 可能会出现涂抹带甚至无特异性条带。
c. 用移液器轻轻吹打混匀或轻微 Vortex 混匀,室温离心数秒,使液体积聚于管底。
d. 各设置好的 PCR 反应管置于 PCR 仪上,开始 PCR 反应。
2. PCR 反应参数的设置:
本产品是经过适体修饰型 Hotstart Taq DNA Polymerase High-concentration,为了使 PCR 的扩增效率、定量精度等达到最佳,建议热激时间为 95℃ ,10 分钟
3.产品包装:
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Product description:
Hotstart Taq DNA Polymerase High concentration is a thermostable TAQ DNA polymerase modified by an aptamer group. Before high temperature heating, the aptamer group combines with Taq enzyme to inhibit the activity of polymerase, avoid non-specific amplification of primer extension or the production of primer dimer, enhance the specificity, sensitivity and stability of DNA amplification, and can be widely used in conventional PCR, multiple PCR, nested PCR, etc. After heat shock at 95 ℃ for 10 min, the enzyme can recover its activity. In addition, Hotstart Taq DNA Polymerase High concentration did not detect 3 '→ 5' exonase activity, but it has 5 '→ 3' exonase activity, which can be used for fluorescent quantitative PCR detection. Hotstart Taq DNA Polymerase High concentration has no activity at room temperature, which is convenient for the normal temperature operation of PCR experiment.
Product content:
1. Hotstart Taq DNA Polymerase High-concentration (10 U/ μ l )
2. 5 × Hotstart Taq Buffer ( Mg²⁺ Plus)
3. Solution I (10 ×)
Activity definition: at 74 ℃ for 30 min, the amount of enzyme required for 10 nmol dNTP to be mixed into acid insoluble sediment is defined as one activity unit.
usage method:
1. Setting of PCR reaction system:
a. Dissolve and mix all solutions required for PCR reaction. It shall be placed on the ice bath or in the ice box. It is recommended that reaction PCR liquid be used separately to avoid repeated freezing and thawing.
b. Refer to the following table to set up PCR reaction. It is recommended that the PCR reaction system be configured in an ice bath or on an ice box:
※ Dosage of template DNA: To ensure the sensitivity of reaction, 25 μ L The system uses the target sequence copied from 10⁴ as the template. Please refer to the following table to calculate the template amount to be added to the PCR system.
one μg Moles of DNA from various sources
For example, the concentration of purified human genome DNA is 1 μ g/ μ l. The number of copies of a gene in the human genome is 10, and the number of copies per unit volume is:
3.0× 10⁵ mol/μg × 1 μg/μl × 10 copy/mol
=3.0× 10⁶ copy/μl
1× 10⁴ copy/ (3.0× 10⁶ copy/μl)
= 1/300 μl
That is, the concentration of 1/300 ul is 1 ug/ μ The human genome DNA of L contains 10⁴ copies of this gene, diluted 300 times and then added 1 μ L to 25 μ L PCR system. To ensure the specificity of the reaction, the final concentration of DNA should be less than 10 ng/ul, and excessive DNA may have smear bands or even no specific bands.
c. Use a pipette to gently blow and mix or slightly Vortex and centrifuge at room temperature for several seconds to make the liquid volume concentrate at the bottom of the tube.
d. Place each set PCR reaction tube on the PCR instrument to start PCR reaction.
2. Setting of PCR reaction parameters:
This product is a aptamer modified Hotstart Taq DNA Polymerase High concentration. In order to optimize the amplification efficiency and quantitative accuracy of PCR, it is recommended that the heat shock time be 95 ℃, 10 minutes
3. Product packaging:
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通过匹配包装上的批号来查找并下载产品的 COA,每批产品都进行了严格的验证,您可放心使用!
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | H292167 | |
| 分析证书 | H292167 |