Biotechnology
Biology 122
Genes and Development
Restriction enzymes
←Produces blunt ends
Red arrows are the cut sites; Green dots indicate the center of the sequence
Restriction
analysis
Cloning into plasmids
Production of
cDNA
Colony
hybridization
Southerns
RFLP analysis
PCR
Dideoxynucleotides
Manual DNA
sequencing
Sequencing by synthesis
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Adapter
DNA fragment
Dense primer lawn
in flow cell
Adapter
DNA
Adapters
Flow cell
a.
1 cm
b.
b: © 2007, Illumina Inc. All rights reserved
Animation of Illumina sequencing
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Bridge
amplification
with unlabeled
dNTPs
c.
Free end
binds to
primer
Fragments
become
doublestranded
d.
Attached
Free
terminus
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Bridge
amplification
with unlabeled
dNTPs
c.
Free end
binds to
primer
Fragments
become
doublestranded
d.
Attached
Free
terminus
Denature
doublestranded
molecules
e.
Attached
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Bridge
amplification
with unlabeled
dNTPs
c.
Free end
binds to
primer
Fragments
become
doublestranded
d.
Attached
Free
terminus
Denature
doublestranded
molecules
e.
Attached
35 cycles
of bridge
amplification
f.
Clusters
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
G
C
T
A
T
G
C
A
First round of
synthesis with
labeled dNTPs
g.
Yeast
Two-hybrid
system
Construction of knockout mice
Construction of knockout mice
Normal
mouse
HGH
mouse
Transgenic mouse
Ventral view of a 14-dayold transgenic mouse,
carrying a reporter gene
that is active in the
developing limb.
Stereomicroscopy Photo by
Dr. Mohammad K. Hajihosseini
University of East Anglia, Norwich, U.K.
Bioscapes 2009 photo contest
Gene therapy
Insertion of genes using the Ti plasmid
Genes inserted into rice to produce Golden rice