1 M Dithiothreitol (DTT)

1 M二硫苏糖醇（DTT）

Company: Thermo Fisher Scientific

Catalog#: P2375

Cytopathic Effect Assay and Plaque Assay to Evaluate in vitro Activity of Antiviral Compounds Against Human Coronaviruses 229E, OC43, and NL63

Yanmei Hu, Chunlong Ma and Jun Wang

Published online: 2022-02-05

Bio-protocol()

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RNase H Polymerase-independent Cleavage Assay for Evaluation of RNase H Activity of Reverse Transcriptase Enzymes

Author:

Angela Corona and Enzo Tramontano,

Date:

2015-08-20

[Abstract] The ribonuclease H (RNase H) polymerase-independent cleavage assay allows detection and quantification of RNase H activity of reverse transcriptase (RT) enzymes with a hybrid substrate formed by a fluorescein labeled RNA annealed with Dabcyl DNA (Figure 1). Here we describe a protocol that we have adapted for HIV-1 RT expressed from a p(His)6-tagged p66/p51 HIV-1HXB2 RT-prot plasmid and for RT of the prototype foamy virus (PFV RT).

Figure 1. Scheme of the principle of the experiment. The RNA substrate (blue) labeled with the fluorophore fluorescein (F, yellow) is annealed with complementary DNA strand (green) labeled with a quencher molecule Dabcyl (D, red). Panel ... [摘要] 核糖核酸酶H（RNase H）聚合酶非依赖性切割测定允许用由与Dabcyl DNA退火的荧光素标记的RNA形成的杂交底物检测和定量逆转录酶（RT）酶的RNA酶H活性（图1）。在这里我们描述了一个协议，我们已适应的艾滋病毒1 RT表示从p（His）6标记的p66/p51艾滋病毒1 HXB2 RT-prot质粒和原型泡沫病毒（PFV RT）RT。

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1 M Dithiothreitol (DTT)

1 M二硫苏糖醇（DTT）

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