Measuring the Activity of Φ29 DNA Polymerase

The major goal of this large project is to develop a Recognition Tunneling Nanopore (RTP) device that will be used for determining the structure of glycosaminoglycans (GAGs). The RTP device is composed of a recognition tunneling junction that is embedded in a nanopore. In order to translocate the GAG molecule through the nanopore, researchers have designed a scheme in which the GAG molecule of interest will be attached to the 5’ end of a DNA primer (figure 1) and the DNA primer will be extended by a biotinylated Φ29 DNA polymerase that is anchored in the nanoslit using streptavidin. This research project specifically is part of a larger project with the main goal of comparing the activity of the wild-type Φ29 DNA polymerase which I have expressed and purified with the mutated Φ29 DNA polymerase devoid of 3’ - 5’ exonuclease activity which was made by Dr. Deng.