Download Slide 1 - Loyola Blakefield

BIOTECH!
Figure 20.17 DNA fingerprints from a murder case
Figure 20.0 DNA sequencers
Biotechnology
• Biotechnology is the manipulation of
organisms or their components to provide
useful products or perform practical tasks
DNA Cloning
• Produces gene-sized pieces of DNA in
multiple identical copies.
• Plasmids, circular DNA pieces separate
from the main chromosome, are used
• Human growth hormone is mass-produced
this way
Figure 20.1 An overview of how bacterial plasmids are used to clone genes
Restriction Enzymes
• Cut up DNA into fragments after a certain
base sequence is recognized
Figure 20.2 Using a restriction enzyme and DNA ligase to make recombinant DNA
Figure 20.4 Using a nucleic acid probe to identify a cloned gene
Polymerase Chain Reaction
• Also called PCR.
• Method of copying small samples of DNA
multiple times. Involves heating of DNA
strands (denaturing) and cooling.
Nucleotides and primers in solution initiate
replication.
Figure 20.7 The polymerase chain reaction (PCR)
Gel Electrophoresis
• Used to separate small DNA fragments
(typically treated first with restriction
enzymes) on the basis of their mass and
charge
• Heavier fragments travel more slowly
down the gel than lighter fragments
Figure 20.8 Gel electrophoresis of macromolecules
Figure 20.9 Using restriction fragment patterns to distinguish DNA from different
alleles
Southern Blotting
• Enables researchers to detect whether
certain DNA sequences exist in a sample.
• Bands from electrophoresis are “blotted”
onto a special paper, and treated with a
radioactive DNA single strand.
• The radioactive strand binds with any
complementary strand
Figure 20.10 Restriction fragment analysis by Southern blotting
Sanger Method
• Used to determine the nucleotide
sequence of DNA.
• DNA fragments and primers, and special
modified nucleotides (dd) used.
• DNA sequence can be read off of gel.
Figure 20.0 DNA sequencers
Figure 20.12 Sequencing of DNA by the Sanger method (Layer 1)
Figure 20.12 Sequencing of DNA by the Sanger method (Layer 2)
Figure 20.12 Sequencing of DNA by the Sanger method (Layer 3)
Figure 20.12 Sequencing of DNA by the Sanger method (Layer 4)
Figure 20.12 Sequencing of DNA by the Sanger method (Layer 4)
ddATP
ddCTP
ddTTP
ddGTP
Figure 20.12 Sequencing of DNA by the Sanger method (Layer 4)
ddATP
ddCTP
ddTTP
ddGTP
Nucleotide Sequence:
ATCAGTAGTGCT
Original DNA Sequence: TAGTCATCACGA
E
E
Table 20.1 Genome Sizes and Numbers of Genes
Figure 20.x1a Laboratory worker reviewing DNA band pattern
Macroorganism Cloning
• Nucleus of an adult cell is removed,
placed into an egg cell whose nucleus was
removed.
• Egg cell is stimulated to start dividing.
• Organism is a clone of its mother.