Size Matters: DNA Amplification

Overview

Objectives/Goals To optimize the PCR process and sequence my successfully amplified DNA. Methods/Materials PCR was performed with standard reagents and thermal cycling parameters. MgCl(2)concentration was varied, in addition to the annealing temperature, and agarose gel staining technique. DNA was sequenced and edited. Results In this experiment, the highest MgCl(2)concentration (8ul) and the higher annealing temperature (48 degrees C)produced the most amplified DNA. In addition, the ethidium bromide gel staining technique worked dramatically better than Ward's Quick View DNA stain. Cytochrome b gene fragment sequences were obtained for two individuals of the intertidal fish, Clinocottus analis. Conclusions/Discussion These results indicate that the highest MgCl(2) concentration improved the enzynmatic activity of the DNA polymerase. The increased annealing temperature allowed the primer to bind to the right complementary bases of the template DNA strand. The ethidium bromide DNA stain illuminated the amplified DNA better because the technique required ultra-violet light that made it easy to visualize the amplified DNA.

Competition history

• CSEF 2005 Biochemistry/ Molecular Biology · Entry S0406

Resources

• CSEF project summary ↗

Related projects

Closest projects by meaning, across every fair and year in the corpus.

Browse more like this

• All Cellular and Molecular Biology projects
• CSEF Cellular and Molecular Biology projects
• Every CSEF 2005 project