16 rflp analysis-2010 update

1,361 views
1,067 views

Published on

Published in: Technology, Education
0 Comments
0 Likes
Statistics
Notes
  • Be the first to comment

  • Be the first to like this

No Downloads
Views
Total views
1,361
On SlideShare
0
From Embeds
0
Number of Embeds
2
Actions
Shares
0
Downloads
0
Comments
0
Likes
0
Embeds 0
No embeds

No notes for slide

16 rflp analysis-2010 update

  1. 2. RFLP Analysis <ul><li>restriction fragment length polymorphism </li></ul><ul><li>differences in DNA sequence on homologous chromosomes (non-coding regions) that result in different DNA fragments when cut by a restriction enzyme (look at your handout). </li></ul><ul><li>polymorphism – differences between DNA sequences </li></ul><ul><li>(e.g. alleles) </li></ul><ul><li>Individuals can be differentiated by slight changes in DNA sequences. </li></ul><ul><li>http://highered.mcgraw-hill.com/olc/dl/120078/bio20.swf </li></ul>
  2. 3. Polymorphism example
  3. 4. RFLP Analysis <ul><li>Individuals can be identified by looking at the polymorphisms </li></ul><ul><li>method only works when the sequence you are looking for contains the restriction site </li></ul>
  4. 7. Large genomes cut with REs will look like a smear after electrophoresis. Each lane contains DNA from a different individual.
  5. 8. Southern Blotting <ul><li>Southern blot – transfer of gel electrophoresis DNA to a nylon membrane </li></ul><ul><li>(no electricity is required!) </li></ul><ul><li>Membrane can be used over and over again for multiple analysis. </li></ul>
  6. 9. Southern Blot nylon membrane electrophoresis gel DNA transferred to membrane
  7. 10. Analysis Probe <ul><li>RFLP probe – specific (generic DNA), radioactive DNA sequences are exposed to the membrane </li></ul><ul><li>(e.g. any known disease) </li></ul><ul><ul><li>nylon membrane is placed </li></ul></ul><ul><ul><li>in a bag with liquid </li></ul></ul><ul><ul><li>containing radioactive DNA </li></ul></ul>
  8. 11. RFLP Analysis <ul><li>method only works when the sequence you are looking for contains the restriction site </li></ul><ul><li>radioactive DNA (generic DNA) only binds to membrane where the sequences are a complementary match </li></ul>
  9. 12. RFLP Autoradiograph <ul><li>photographic film used to detect the sequence </li></ul>
  10. 13. http://www.biology.arizona.edu/Human_bio/activities/blackett/introduction.html
  11. 15. control lanes indicate that the samples were prepared properly
  12. 17. Who’s related to Fred and Norma?
  13. 20. Semen Analysis

×