MCDB 2150 Lecture 3: Class 3 DNA Visualization and Forensics

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DNA Visualization and Forensics
Class Notes:
PCR recap:
o PCR makes millions of copies of a specific sequence, designated by primers
o Steps:
Denature DNA (95 degrees C)
Anneal Primers (65 degrees C)
Extend Primers (75 degrees C)
Repeat
DNA Visualization Gel electrophoresis
o Gel electrophoresis separates DNA on the basis of molecular weight
(size)
o Pieces of DNA amplified through PCR can be visualized with a dye that
intercalates between strands of DNA and fluoresces under UV light
o The gel is like a mesh material
o Load DNA at one side: DNA is negatively charged
o Run electrical current through the material
When a current is run through the system: the short pieces of
DNA will move farther towards the positive electrode
o All pieces are negatively charged and move away from the negative
electrode
o Big pieces of DNA can’t move through the gel as quickly (more friction)
o Smaller pieces of DNA can move through quickly (less friction)
o A cartoon of a gel looks like this:
each band represents all the DNA molecules in the sample that are a particular
size
if a band is thicker, that means there is more DNA molecules of that size
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