We have examined the replication patterns of ribosomal DNA plasmids in vivo and in vitro using Xenopus eggs. Plasmids carrying different parts of the Xenopus ribosomal DNA sequence were allowed to replicate either in vitro in an egg extract or in vivo after microinjection into unfertilized eggs. The replication intermediates were analyzed by the 2D gel electrophoretic technique of Brewer and Fa...

Isoelectric focusing (IEF) is the first step for two-dimensional (2D) gel electrophoresis and plays an important role in sample purification for proteomics. However, biases in protein size and pI resolution, as well as limitations in sample volume, gel capacity, sample loss, and experimental time, remain challenges. In order to address some of the limitations of traditional IEF, we present a mi...

Two-dimensional (2D) gel electrophoresis allows separation of complex mixtures of proteins based on isoelectric points and relative mobility. This method has not changed much fundamentally since their original description in the late 1970s. Despite several limitations, such as solubilization of membrane proteins and separation of highly basic proteins, this method has been used successfully in ...

We describe an algorithm, Contour Area Filtering, for separating background from foreground in gray scale images. The algorithm is based on the area contained within gray scale contour lines. It can be viewed as a form of local thresholding, or as a seed growing algorithm, or as a type of watershed segmentation. The most important feature of the algorithm is that it uses object area to determin...

4) Briefly describe any new skills you acquired during your summer research: Over the course of the summer, I have learned to optimize many facets of the gel electrophoresis method. I have optimized the construction of a low cost electrophoresis chamber, various tendon cutting techniques, run times, voltage levels, electrode material, and gel analysis techniques. I have also been taught to opti...