1 / 13

Gel Electrophoresis

Lorena Gomez 05 November 2014. Gel Electrophoresis. Overview. What is gel electrophoresis How does is work? What is it good for? Who uses it Example applications DGGE (forensics) SCGE (microbiology). Gel Electrophoresis.

jaden-valenzuela
Download Presentation

Gel Electrophoresis

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Lorena Gomez 05 November 2014 Gel Electrophoresis

  2. Overview • What is gel electrophoresis • How does is work? • What is it good for? • Who uses it • Example applications • DGGE (forensics) • SCGE (microbiology)

  3. Gel Electrophoresis • Method of separating and analyzing micromolecules and their fragments • DNA, RNA, Proteins • Size • Charge

  4. Gel Electrophoresis

  5. DNA/RNA vs Protein • DNA and RNA are negatively charged and will be attracted to the positive electrode and migrate down. However, proteins are positively charged, this means that before being able to use this technique the protein must be negatively coated allowing the protein to migrate downwards.

  6. Dying • After the electrophoresis is complete, the sample has to be stain in order to see the results. There are different dyes available for staining depending on what macromolecule is being observed. • Examples • Sodium dodecylsulfate • Ethidium Bromide • Silver stain

  7. Field Applications • Biochemistry • Microbiology • Molecular biology • Forensics • Genetics

  8. “Analysis of postmortem DNA degradation by single cell gel electrophoresis” • By Laura A Johnson • In Johnson’s paper, she goes on to explain the importance of determining the time of death of a victim in forensic science • Johnson looks into the possibility of using single cell gel electrophoresis (SCGE) to study postmortem cell processes. • Fragmintation after death • Johnson used this application on a human blood (leukocyte) model.

  9. Summery of Johnsons Paper • Johnsons results showed an increase in the DNA fragmentation from 2 to 22 h after removal from the body. • After the 56 h point the image does not really change. • The DNA fragmentation proved to be dependent on the organ examined and on time

  10. “Detention and Identification of Gastrointestinal Lactobacillus Species by Denaturing Gradient Gel Electrophoresis and Species-Specific PCR Primers” • By J. Walter. • Uses denaturing gradient gel electrophoresis (DGGE) to detect the presence of Lactobacillus species in the stomach content of mice. • Walter proposes that using a molecular method along with the traditional once would give results that are more reliable.

  11. Summery of Walter’s Paper • Using the 16s nRNA gene sequence allowed for modern bacteria classification, thus DGGE along with PCR primers (that target this gene) could allow for a more practical approach to identifying lactobacillus. • Using DGGE he was able to better detect the content and changes in the microbial community

  12. Summery of Walter’s Paper • Using PCR-DGGE showed to have potential in finding species that could be missed using traditional methods

  13. Bibliography • Gel Electrophoresis. Amsterdam: Elsevier, 1995. N. pag. Biological Sciences Initiative. Web. 02 Nov. 2014. <http://www.colorado.edu/Outreach/BSI/pdfs/gel_electrophoresis.pdf>. • "Gel Electrophoresis." Gel Electrophoresis. Genetic Science Learning Center, n.d. Web. 02 Nov. 2014. <http://learn.genetics.utah.edu/content/labs/gel/>. • "Gel Electrophoresis." Nature.com. Nature Publishing Group, n.d. Web. 02 Nov. 2014. <http://www.nature.com/scitable/definition/gel-electrophoresis-286>. • . Walter, G. W. Tannock, A. Tilsala-Timisjarvi, S. Rodtong, D. M. Loach, K. Munro, and T. Alatossava. Appl. Environ. Microbiol. January 2000 66:1 297-303; doi:10.1128/AEM.66.1.297-303.2000 • L. Johnson, J. Ferris, Analysis of postmortem DNA degradation by single-cell gel electrophoresis, Forensic Science International, Volume 126, Issue 1, 28 March 2002, Pages 43-47, ISSN 0379-0738, http://dx.doi.org/10.1016/S0379-0738(02)00027-0.

More Related