PCR within Powdered Porous Glass: Potential Application for Glass-coated Amorphous Metal Microwires
Date
2004
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Alfred University. Faculty of Ceramic Engineering. Kazuo Inamori School of Engineering.
Abstract
The polymearse chain reaction (PCR) has become one of the most powerful techniques in biology. The aim of this sutdy was to take the first step towards the development of an in situ PCR device with porous glass-coated amorphous metal microwires. In an attempt to run PCR encased within a porous glass structure, prototypes were to be developed such that they could hold an entire 50μL reation. The glass composition, 6.5Na_{2}O*33.5B_{2}O_{3}*60.0SiO_{2} (mol%), was heat-treated at 680ºC for 28 min to develop phase separation on the order of of 100nm. This glass could not be leached in 55 wt% H_{2}SO_{4} without cracking. To prevent fracture, a vapor leach method was developed to leach samples near their annealing temperature. This technique could not form porous glass subjects with deep enough pores. Instead, powdered Controlled Pore Glass® (CPG®) was obtained from Millipore Corporation for use in PCR experimentation. DNA amplification in the presence of powdered Uncoated CPG® was unsuccessful due to DNA polymerase adsorption. This problem could not be counteracted with competitive binding of bovine serum albumin. CPG® Glyceryl (CPG®-G) was used for PCR experimentation instead. This glass was made with a glycerol-derived coating. DNA amplification was successful with reactions completely aborbed by powdered CPG®-G.
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Keywords
Coatings, Porous glasses, PCR, Microwires, Amorphous metals, DNA, Enzymes