| Assay Method Information | |
| | HIV-1 RT Inhibition Assay |
| Description: | A reverse transcriptase (RT) assay kit produced by Roche was selected for the RT inhibition assay. All the reagents for performing the RT reaction came with the kit, and the ELISA procedures for RT inhibition assay were carried out following the description in the kit protocol. Briefly, the reaction mixture containing template/primer complex, viral nucleotides (dNTPs), and RT in the incubation buffer with or without inhibitors was incubated for 1 h at 37 °C. After that, the reaction mixture was transferred to a streptavidincoated microtiter plate and incubated for another 1 h at 37 °C to make sure that retranscriptional cDNA chain consisted of biotin-labeled dNTPs bound to streptavidin. Then, unbound dNTPs were removed using washing buffer and anti-DIG-POD working solution was added. After incubation for 1 h at 37 °C, the DIG-labeled dNTPs incorporated into cDNA were bound to the anti-DIG-POD antibody. The unbound anti-DIG-PODs were removed. |
| Affinity data for this assay | |
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