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DNA Profiling and Gel Electrophoresis
Biology Accelerated 2006
Biotechnology Genetics
Some slides taken and edited from Maria Hall, Julie Basil and Karen Wolfe
Commonly Used Techniques
Each of the following will be explained in further detail Restriction enzymes Recombinant DNA (using plasmids) Gel electrophoresis DNA profiling DNA sequencing
Restriction Enzymes “molecular scissors”
Enzymes found in bacteria Used by the bacterium as a defense against
“foreign” DNA Cut DNA at very specific places or
sequences (called restriction sites) The “cut” DNA pieces can have either
Blunt ends Sticky ends
Restriction Enzymes
also known as restriction endonucleases
named for the bacteria where they originally came from ex: EcoRI
Why cut DNA?
DNA is cut so that another piece of DNA can be inserted
The “other” DNA contains the gene you want to copy
Citation Greatest Discoveries with Bill Nye: Genetics. Discovery Channel School. 2005. unitedstreaming. 23 January 2006 <http://www.unitedstreaming.com/>
Citation (MLA)Biologix: Manipulating DNA. United Learning. 1997. unitedstreaming. 23 January 2006 <http://www.unitedstreaming.com/>
cut each DNA strand at different bases--creates “sticky ends”
open “flap” of DNA can readily bind with a complementary “flap”
some restriction enzymes make “blunt ends”
RFLPs
abbreviation pronounced “rifflips” stands for restriction fragment length
polymorphisms restriction fragment =
fragment of DNA cut with restriction enzymes
length polymorphisms = many different lengths
Gel electrophoresis
technique used to separate strands of DNA, RNA or protein according to their size
For DNA, samples first need to be purified and cut with restriction enzymes
DNA placed into a well at one end of an agarose gel (like jello)
the gel acts as a strainer
Citation Greatest Discoveries with Bill Nye: Genetics. Discovery Channel School. 2005. unitedstreaming. 23 January 2006 <http://www.unitedstreaming.com/>
Electrophoresis animation
Electrophoresis Animation Electrophoresis Animation 2
http://www.bbc.co.uk/education/asguru/biology/04genesgenetics/04geneticcode/01geneticengineering/02isolatinggenes/index.shtml
http://www.shsu.edu/~chm_tgc/sounds/flashfiles/GE.swf
Self Check and Review
In your study group of 3 students discuss the following issues form a consensus and write down what you learned: DNA Profiling
Why is it done? What are its uses? What is needed to do DNA profiling (materials). How is it done? (steps in the process)
PCR ClipCitation (MLA)Biologix: Manipulating DNA. United Learning. 1997. unitedstreaming. 23 January 2006 <http://www.unitedstreaming.com/>