Assay Method Information

Assay Name:  HIV-1 RT In-Vitro Assay
Description:  The reaction mixture was set with template primer complex, RT enzyme and dNTPs in a lysis buffer with or without inhibitors. The reaction mixture was incubated at 37 °C for 1 h and then transferred to streptavidine-coated microtitre plate (MTP). The biotin-labeled dNTPs that were incorporated in the template due to activity of RT, bound to streptavidine. The unbound dNTPs were washed using wash buffer and anti-DIG-POD was added to the MTP. The DIG-labeled dNTPs incorporated in the template were bound to an anti-DIG-POD antibody. The unbound anti-DIG-POD was washed again with washing buffer and the peroxide substrate (ABST) was added to the MTP. A colored reaction product was produced during the cleavage of the substrate catalyzed by a peroxide enzyme. The absorbance of the sample was determined as an optical density (OD) at 405 nm using a micro titer plate ELISA reader.
Affinity data for this assay
 

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