Real-Time Fluorescence Assays to Monitor DNA Polymerase Activities

Quantitative Real-time Polymerase Chain Reaction (qRT-PCR). Fatemeh Mehrpooyan*

Single-molecule and ensemble fluorescence assays for a functionally important conformational change in T7 DNA polymerase.

We report fluorescence assays for a functionally important conformational change in bacteriophage T7 DNA polymerase (T7 pol) that use the environmental sensitivity of a Cy3 dye attached to a DNA substrate. An increase in fluorescence intensity of Cy3 is observed at the single-molecule level, reflecting a conformational change within the T7 pol ternary complex upon binding of a dNTP substrate. T...

John Eid , Molecules Real - Time DNA Sequencing from Single Polymerase

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Real-time DNA sequencing from single polymerase molecules.

Pacific Biosciences has developed a method for real-time sequencing of single DNA molecules (Eid et al., 2009), with intrinsic sequencing rates of several bases per second and read lengths into the kilobase range. Conceptually, this sequencing approach is based on eavesdropping on the activity of DNA polymerase carrying out template-directed DNA polymerization. Performed in a highly parallel op...

Molecules Real - Time DNA Sequencing from Single Polymerase

http://www.sciencemag.org/cgi/content/full/323/5910/133 version of this article at: including high-resolution figures, can be found in the online Updated information and services, http://www.sciencemag.org/cgi/content/full/1162986/DC1 can be found at: Supporting Online Material found at: can be related to this article A list of selected additional articles on the Science Web sites http://www.sc...