Transcript BIO-RAD_DNA_fingerprinting

BIO-RAD Forensic DNA
Fingerprinting Kit
Section Concepts: DNA Scissors
Activity, restriction enzymes, gel
electrophoresis, standard curve
Background Information
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Restriction enzymes:
found in bacteria and
archaea, are enzymes
that cuts DNA at
specific recognition
nucleotide sequences
known as restriction
sites.
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Gel Electrophoresis: a
method used in
biochemistry and
molecular biology to
separate a mixed
population of DNA and
RNA fragments by
length, to estimate the
size of DNA and RNA
fragments, or to
separate proteins by
charge. Nucleic acid
molecules are
separated by applying
an electric field to move
the negatively charged
molecules through an
agarose matrix.
Restriction Enzymes + Gel
Electrophoresis
Restriction enzymes video
Gel Electrophoresis animation
Timeline/Overview
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Monday: Lecture
Tuesday: Activities 1 & 2, DNA scissors,
pre-lab quiz
Thursday: DNA fingerprint (restriction
digest), Analyze lab (work on packets)
Monday: No school
Tuesday: DNA fingerprinting kit (run gel,
visualize)
Thursday: Packet due, post quiz
Necessary Preparation for
Lab DNA Finger printing
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DNA fingerprinting
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Labels
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8—clear 1.5mL tubes ENZ (enzyme)
8—clear 1.5mL tubes M (marker)
8—clear 1.5mL tubes LD (loading dye)
8--green 1.5mL tubes CS
8—blue 1.5mL tubes S1
8—orange1.5mL tubes S2
8—violet 1.5mL tubes S3
8—red 1.5mL tubes S4
8—yellow 1.5mL tubes S5
Before Digest
• Set water bath for 37C
• Rehydrate & aliquot (page 13)
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EcoRI/PstI enzyme mix (12 hours before use)
Crime scene DNA
Suspects 1-5 DNA
Before Running Gel
• Prepare electrophoresis TAE buffer
• 11/13 at before 5th period Pour 8 gels (1%, EtBr, 10 well comb)
• Prep. HindIII standard (right before class)
• Aliquot
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HindIII (marker)
Loading dye
DNA Fingerprinting Lab Background Information
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DNA
Fingerprinting:
technique used by
scientists to
identify an
individual based
on their specific
DNA profile
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Restriction
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Digestion of
DNA: restriction
enzymes
recognize
particular
sequences in a
segment of DNA
and cuts the DNA
molecule at that
point
Plasmids:
circular, nonchromosomal
pieces of DNA
that can replicate
in and are
commonly found
in bacteria
DNA Fingerprinting
DNA Fingerprinting Video
Plasmids
Restriction Enzyme Digestion
of Plasmid DNA
Restriction Enzyme
Animation
Restriction Enzyme
EcoR1 Animation
DNA Fingerprinting Tips
Place tubes in refrigerator after
removing from water bath
 Gel will be 10 wells, adjust lanes in
your protocol to use 2-8
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