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文献和实验A Method to Assay Inhibitors of DNA Polymerase IIIC Activity
, one of these, designated pol III, is responsible for DNA replication and growth of bacteria and, therefore, warrants consideration as a drug target. However, the pol III enzymes of Gram-positive and Gram-negative species are quite different, and the Gram-positive enzyme
Enzyme Kinetics assay of the WT
To assay 17 b-HSD activity in lysates, cells were harvested 48h after transfection using PBS Enzyme Free Cell Dissociation Solution ( specialty Media Inc., Lavellette, NJ), frozen on dry ice and stored at -80 C. Cell pellets were thawed
Cloning Polymerase Chain Reaction Products Utilizing theT/A Overhang and a Kit
to the ligation reaction mixture. This procedure is based on the fact that Taq polymerase has a nontemplate dependent activity that adds a single deoxyadenosine to the 3′ ends of double-stranded DNA (2 ). The Invitrogen pCR™II vector is supplied as a linearized
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