Lecture 7 Recombinant DNA technology III Electrophoresis and sequencing

Lecture 7 Recombinant DNA technology III Electrophoresis and sequencing

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Biol142 Foundations in Modern Biology II Cellular Biology and Genetics Lecture 7 – Wednesday, February 1 st , 2012 Recombinant DNA technology III: Gel electrophoresis and DNA sequencing. Reading: Freeman, Emory 2 nd Ed. Chapter 19 (pages 346-347)
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Learning objectives At the end of this lecture, you will be able to: 1) Predict the migration of DNA fragments of differing size on an agarose gel. 2) Use a DNA ladder to determine whether a plamid contains an additional DNA insert. 3) Describe the use of Dideoxynucleotide triphosphates (ddNTPs) in DNA sequencing reactions.
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1. How can we isolate DNA fragments of interest? DNA fragments can be separated using gel electrophoresis . This method takes advantage of the negative charge of DNA at neutral pH. G el electrophoresis involves: 1. Adding DNA to a semisolid gel. 2. Application of an electric field to the gel. 3. The negatively charged DNA molecules migrate toward the positive pole. Smaller molecules can migrate more quickly through the porous gel than larger ones. Small fragments migrate faster toward +ve pole.
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stained with a fluorescent dye and examined under ultraviolet light. What does
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Lecture 7 Recombinant DNA technology III Electrophoresis and sequencing

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