Lab # 9 : Restriction enzyme mapping Restriction Enzymes Restriction - PowerPoint PPT Presentation

Jan 31, 2023 •137 likes •229 views

Lab # 9 : Restriction enzyme mapping Restriction Enzymes Restriction enzymes , also known as restriction endonucleases, are enzymes that cut a DNA molecule at a particular place (=recognition site). A blunt end : A sticky end:

Lab # 9 : Restriction enzyme mapping
Restriction Enzymes Restriction enzymes , also known as restriction endonucleases, are enzymes that cut a DNA molecule at a particular place (=recognition site). A blunt end : A sticky end: https://askabiologist.asu.edu/restriction-enzymes http://www.shmoop.com/biotechnology/dna-technology.html
Restriction Enzyme Mapping Kojic et al. 2011, BMC Microbiology https://en.wikipedia.org/wiki/File:PBR322_plasmid_showing_restriction_sites_and_resistance_genes.jpg
Today’s lab Each pair does 4 reaction tubes
Gel Electrophoresis _ + Run to the red! DNA is strongly negative => it will migrate through the gel towards the positive electrode (from cathode to anode) http://tymkrs.tumblr.com/post/3069263517/gel-electrophoresis-negative-to-positive
DNA migration in Agarose Gel • The pores in the gel separate the DNA molecules according to their size and shape. • The smaller the DNA molecule, the faster it migrates through the gel. Annals of Neurosciences https://passel.unl.edu/pages/printinformationmodule.php?idinformationmodule=1065724861
Visualization of DNA migration Ladder The tracking dye moves along with the DNA sample Ladder Two dyes can be used https://passel.unl.edu/pages/printinformationmodule.php?idinformationmodule=1065724861 http://tymkrs.tumblr.com/post/3069263517/gel-electrophoresis-negative-to-positive
Types of DNA Molecules linear supercoiled circular • supercoiled DNA migrates faster than its linear form • linear DNA migrates faster than its nicked circular form http://laneccgenetics.pbworks.com/w/page/58169624/Chromosomes%20Overview https://pixabay.com/en/photos/dna/ amanaimages.com http://www.bioch.ox.ac.uk/aspsite/index.asp?pageid=1118

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