Understanding DNA Fragment Analysis Through Polyacrylamide Gel Electrophoresis
Polyacrylamide gel electrophoresis is a technique used to separate DNA fragments based on size. By casting a gel and running an electric current through it, terminated DNA fragments can be separated and visualized using isotopes. This process allows for the analysis of DNA fragments differing in size by just one base pair. Techniques such as linear PCR and cycle sequencing in the 1980s paved the way for more efficient DNA amplification methods.
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After synthesis, a polyacrylamide gel was cast Acrylamide is neurotoxic until polymerized
The terminated fragments are separated according to size on the gel. After electrophoresis, the gel is dried and exposed to photographic film. The decay of Isotope-labeled dCTP (filled circles) produces photons, that blacken the film. PAGE allows us to distinguish DNA fragments that differ in size by only one base pair large small
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1980s: Linear PCR is used to generate more products from less template Cycle sequencing is a simple method in which successive rounds of denaturation, annealing, and extension in a thermal cycler result in linear amplification of extension products. From applied biosystems.com
