User:Klare Lazor/Notebook/Chem-496-001/2011/09/21

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Objective

determine the number of base pairs in the mutated dna

Description

Gel preparation 51 agarose gel

  • 0.01 grams for every mL used
  • 25mL, so 0.25grams of agarose
  • different amounts of base pairs need specific percent of agarose
  • buffer to make use TAE buffer amount 25mL
    • 25mM Tris
    • 190mM Glyox
    • 1%SDS
  • Mix both together does not have to be precise.
  • Microwave for 40 seconds

Data

  • Add data and results here...

Notes

Class Notes

  • Gel electrophoresis background
    • Orginally, DNA fragments were laboriously separated by gravity. Now, they are separated through gel electrophoresis. This process uses electricity to separate DNA fragments by size as they migrate through gel matrix. Gel electrophoresis can be used to separate DNA fragments. The gel is made from purified agarose in powdered form, and is insoluble in water at room temperature. It will dissolve in boiling water. When it cools, it undergoes polymerization. The sugar polymers crosslink with each other; causing the solution to gel into a semi-solid matrix much like jello. The more agarose the firmer the gel will be. The solution is then poured into a casting tray.
    • The technique of electrophoresis is based on the fact that DNA is negatively charge at neutral pH due to its phosphate backbone. As a result, when an electrical potential is place on the DNA it will move toward the positive pole. This causes the DNA strands to be separated by size as they move through the lattice.
    • The DNA must be staine with a fluorescent dye ( usuallly ethidium bromide). Ethidium bromide is very carcinogenic.

http://faculty.plattsburgh.edu/donald.slish/Electrophoresis.html http://www.dnalc.org/resources/animations/gelelectrophoresis.html

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