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NGS Library Prep kits

Molecular Research

Drug Discovery

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T7 Endonuclease I

T7 Endonuclease I recognizes and cuts incompletely matched DNA, cross-shaped structure DNA, Holiday structure or DNA bifurcation points, heterologous DNA; at the same time, it can cut double-stranded DNA with cutting sites at a lower speed. The enzyme cleaves the first, second or third phosphodiester bond at the 5'end of the mismatch site. This product is a high-purity active protein expressed and purified in Escherichia coli after the recombinant T7 endonuclease I gene has been cloned. It can be used for gene mutation, SNP, TALEN or CRISPR/Cas9 mutant detection, identification of mismatched DNA, and decomposition of four Directional cross DNA or branched DNA; detection of heterodimer or cut DNA; or used for random cutting of linear DNA for Shot-gun cloning.   High detection sensitivity; Products can be directly detected by electrophoresis   Store at -20°C
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T7 Endonuclease I recognizes and cuts incompletely matched DNA, cross-shaped structure DNA, Holiday structure or DNA bifurcation points, heterologous DNA; at the same time, it can cut double-stranded DNA with cutting sites at a lower speed. The enzyme cleaves the first, second or third phosphodiester bond at the 5'end of the mismatch site. This product is a high-purity active protein expressed and purified in Escherichia coli after the recombinant T7 endonuclease I gene has been cloned. It can be used for gene mutation, SNP, TALEN or CRISPR/Cas9 mutant detection, identification of mismatched DNA, and decomposition of four Directional cross DNA or branched DNA; detection of heterodimer or cut DNA; or used for random cutting of linear DNA for Shot-gun cloning.

 

High detection sensitivity;
Products can be directly detected by electrophoresis

 

Store at -20°C

Keyword:
T7EndonucleaseI
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Q1:做基因编辑,酶切验证编辑成功,但是测序结果却不正确(假阳性)。

A1:编辑完成之后,提取基因组,做PCR扩增目的片段,利用T7核酸内切酶进行验证,有目的条带,连T载做转化,菌检测序。建议在菌检结束之后,利用菌检产物再做一次酶切验证是否将有效片段连入载体,如果酶切验证是阳性的,则可以测序。

Q2:该实验反应温度的上限是多少?

A2: 反应温度超过42℃时,会增加非特异性核酸酶活性。避免反应温度超过55℃,否则会导致酶活性降低。

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