Restriction enzymes and gel electrophoresis
Download
1 / 8

Restriction Enzymes and Gel Electrophoresis - PowerPoint PPT Presentation


  • 142 Views
  • Uploaded on

Restriction Enzymes and Gel Electrophoresis. Genetics 335. Step 1: Restriction enzymes/endonucleases. Enzymes that cut DNA into fragments of different sizes Cleave DNA at specific phosphodiester bonds Named after bacterial strains E.g., BamHI, EcoRI, HAEIII, HindII, MstII

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about ' Restriction Enzymes and Gel Electrophoresis' - azuka


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.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
Restriction enzymes and gel electrophoresis

Restriction Enzymes and Gel Electrophoresis

Genetics 335


Step 1 restriction enzymes endonucleases
Step 1: Restriction enzymes/endonucleases

  • Enzymes that cut DNA into fragments of different sizes

  • Cleave DNA at specific phosphodiester bonds

  • Named after bacterial strains

    • E.g., BamHI, EcoRI, HAEIII, HindII, MstII

  • Can cleave straight (flush) or staggered

    • Blunt or sticky ends


Why cut up dna
Why cut up DNA?

  • If different alleles can be cut up and identified (using GE), we can identify unknown persons (e.g., criminals, biological relatives), and document unknown genotypes (e.g., Huntington’s Disease)

  • http://www.youtube.com/watch?v=KVnFBCzg8Ms


Gel electrophoresis
Gel Electrophoresis

  • You have a vial of undescribed DNA fragments—now what?

  • Samples pipetted into wells on one end of a gel (e.g., agarose)

  • Electricity is added to the gel

  • DNA fragments move through the gel at different rates, away from the negative and toward the positive end

    • Smaller fragments move easier and further from well, larger fragments more cumbersome and move shorter distances from well

  • The resulting “fingerprint” of fragments can be matched to a known sample or investigated for specific alleles



Videos
Videos

  • http://www.sumanasinc.com/webcontent/animations/content/gelelectrophoresis.html

  • http://www.youtube.com/watch?v=lM8npqHw-mk


Review
Review

  • Restriction enzymes cut up DNA into pieces unique to your DNA

    • DNA pieces are a function of your genetics and the restriction enzyme used

  • DNA fragment soup placed in Gel well and distributes based on fragment/sequence length

  • Resulting gel is unique—like a fingerprint


ad