A high-throughput homogeneous assay for reverse transcriptase using generic reagents and time-resolved fluorescence detection.
A high-throughput homogeneous assay for reverse transcriptase using generic reagents and time-resolved fluorescence detection.
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使用通用试剂和时间分辨荧光检测对逆转录酶进行高通量均相测定。
DOI：
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发表时间：
2000
影响因子：
2.9
通讯作者：
K. Oldenburg
中科院分区：
文献类型：
--
作者：
Ji;Tongming Chen;S. H. Nguyen;K. Oldenburg
A homogeneous time-resolved fluorescence (HTRF) assay has been developed for determining the activity of HIV reverse transcriptase (HIV-RT). By using a sequential capping strategy, the assay has been configured to utilize only generic reagents such as biotinylated dUTP, streptavidin-allophycocyanin, and streptavidin-europium. The assay was optimized for a HIV-RT high-throughput screen. Under optimized conditions, a signal-to-background ratio of approximately 10:1 and a Z' factor of 0.8 were obtained. The titration curves of several known HIV-RT inhibitors have been evaluated with this assay format. This HTRF format can be used for high-throughput assays with other nucleotide polymerases.
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