Biotech

• Recominbinant DNA Technology → think pGLO

• Restriction Enzymes

  • very specific cutting patterns, unique enzyme/pattern for each bacteria

    • blunt cut

    • staggered cut

12/12 Notes for Addai

• Started on Pg7 (Electrophoresis and DNA fingerprinting)

• gel electrophoresis → ability to visualize and separate DNA, RNA, and protein

  • DNA is injected into the wells (rectangular slits) and gentle low is ran through it

  • negative DNA is pushed away from the negative electrode

  • microscopically there’s little holes and nooks in the gel allowing smaller molecules to move through the medium faster →DNA is filtered by size

• DNA fingerprinting is used for

  • Identifying criminals

  • paternity testing

  • mapping the genome

  • diagnostic testing (genetically inherited disease detection)

• DNA fingerprinting is the method to separate the DNA but gel electrophoresis is how it happens

• RFLP (Restriction Fragment Length polymorphism)

  • Test Questions: Using RFLP results for paternity test matching MCQ

  • Digest (cut) large DNA molecules with the restriction enzyme into fragments.

  • Samples containing the fragments are loaded into wells cut into a gel.

  • Electric current in the gel box draws DNA fragments (negatively charged) to the positive pole.

  • As the fragments migrate through the gel they are primarily separated out by size due to the pores in the gel. The shorter fragments move farther because they can travel more easily through the pores (the distance the fragments travel is inversely proportional to their size).

  • The bands of DNA are not visible until they are stained with a dye or identified by a radioactive probe or enzyme-linked tag (Southern blotting). 

• PCR-RFLP Procedure:

  • Remember that PCR just makes millions of copies of a target DNA sequence

• Watch the Race to Sequence the Human Genome