Lecture #9 Date _____

■ Chapter 20~
DNA Technology
& Genomics
O.J. Simpson capital murder case,1/95­9/95
■ Odds of blood in Ford Bronco not being R. Goldman’s:
■ 6.5 billion to 1
■ Odds of blood on socks in bedroom not  being N. Brown­Simpson’s:
■ 8.5 billion to 1
■ Odds of blood on glove not being from R. Goldman, N. Brown­Simpson, and O.J. 
Simpson:
■ 21.5 billion to 1
■ Number of people on planet earth:
■ 6.1 billion
■ Odds of being struck by lightning in the U.S.:
■ 2.8 million to 1
■ Odds of winning the Illinois Big Game lottery:
■ 76 million to 1 
■ Odds of getting killed driving to the gas station to buy a lottery ticket 
■ 4.5 million to 1
■ Odds of seeing 3 albino deer at the same time:
■ 85 million to 1
■ Odds of having quintuplets:
■ 85 million to 1
■ Odds of being struck by a meteorite:
■ 10 trillion to 1
 Recombinant DNA
■ Def: DNA in which genes
from 2 different sources
are linked
■ Genetic engineering:
direct manipulation of
genes for practical
purposes
■ Biotechnology:
manipulation of organisms
or their components to
perform practical tasks or
provide useful products
Bacterial plasmids in gene cloning
 DNA Cloning
■ Restriction enzymes (endonucleases):
in nature, these enzymes protect
bacteria from intruding DNA; they cut up the
DNA (restriction); very specific
■ Restriction site:
recognition sequence for a particular
restriction enzyme
■ Restriction fragments:
segments of DNA cut by restriction
enzymes in a reproducable way
■ Sticky end:
short extensions of restriction
fragments
■ DNA ligase:
enzyme that can join the sticky ends of
DNA fragments
■ Cloning vector:
DNA molecule that can carry foreign
DNA into a cell and replicate there (usually
bacterial plasmids)
Restriction Enzymes

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Steps for eukaryotic gene cloning
■ Isolation of cloning vector
(bacterial plasmid) & gene-
source DNA (gene of interest)
■ Insertion of gene-source DNA
into the cloning vector using the
same restriction enzyme; bind
the fragmented DNA with DNA
ligase
■ Introduction of cloning vector
into cells (transformation by
bacterial cells)
■ Cloning of cells (and foreign
genes)
■ Identification of cell clones
carrying the gene of interest
Cloning

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DNA Analysis & Genomics
■ PCR (polymerase chain
reaction)
■ Gel electrophoresis
■ Restriction fragment
analysis (RFLPs)
■ Southern blotting
■ DNA sequencing
■ Human genome project
Polymerase chain reaction (PCR)
■ Amplification of any
piece of DNA without
cells (in vitro)
■ Materials: heat, DNA
polymerase,
nucleotides, single-
stranded DNA primers
■ Applications: fossils,
forensics, prenatal
diagnosis, etc.
DNA Analysis
■ Gel electrophoresis: separates nucleic acids or proteins on the
basis of size or electrical charge creating DNA bands of the
same length
Restriction fragment analysis
■ Restriction fragment length polymorphisms (RFLPs)
■ Southern blotting: process that reveals sequences and
the RFLPs in a DNA sequence
■ DNA Fingerprinting
 DNA Sequencing
■ Determination of
nucleotide sequences
(Sanger method,
sequencing machine)
■ Genomics: the study of
genomes based on
DNA sequences
■ Human Genome
Project
Practical DNA Technology Uses

■ Diagnosis of disease
■ Human gene therapy
■ Pharmaceutical products
(vaccines)
■ Forensics
■ Animal husbandry
(transgenic organisms)
■ Genetic engineering in
plants
■ Ethical concerns?