Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate...

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Biotechnology Biology 122 Genes and Development

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Transcript of Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate...

Page 1: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Biotechnology

Biology 122

Genes and Development

Page 2: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Red arrows are the cut sites; Green dots indicate the center of the sequence

←Produces blunt ends

Restriction enzymes

Page 3: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 4: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Restriction analysis

Page 5: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 6: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 7: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 8: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Cloning into plasmids

Page 9: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 10: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 11: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 12: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Production ofcDNA

Page 13: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Colonyhybridization

Page 14: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Southerns

Page 15: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

RFLP analysis

Page 16: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 17: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

PCR

Page 18: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Dideoxynucleotides

Page 19: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Manual DNA sequencing

Page 20: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 21: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 22: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

b: © 2007, Illumina Inc. All rights reserved

Adapter

Adapter

Adapters

a. b.

Flow cell1 cm

DNA

DNA fragment

Dense primer lawnin flow cell

Sequencing by synthesis

Animation of Illumina sequencing

http://www.illumina.com/technology/sequencing_technology.ilmn
http://www.illumina.com/technology/sequencing_technology.ilmn
http://www.illumina.com/technology/sequencing_technology.ilmn
Page 23: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

c. d.

Bridgeamplificationwith unlabeleddNTPs

Free endbinds toprimer

Fragmentsbecomedouble-stranded

Freeterminus

Attached

Page 24: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Attached

c. d. e.

Bridgeamplificationwith unlabeleddNTPs

Free endbinds toprimer

Fragmentsbecomedouble-stranded

Freeterminus

Denaturedouble-strandedmolecules

Attached

Page 25: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Attached Clusters

c. d. e. f.

35 cyclesof bridgeamplification

Bridgeamplificationwith unlabeleddNTPs

Free endbinds toprimer

Fragmentsbecomedouble-stranded

Freeterminus

Denaturedouble-strandedmolecules

Attached

Page 26: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

A

A

T

TG

GC

C

g.

First round ofsynthesis withlabeled dNTPs

Page 27: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 28: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Yeast Two-hybridsystem

Page 29: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Construction of knockout mice

Page 30: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Construction of knockout mice

Page 31: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Normalmouse

HGHmouse

Page 32: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Transgenic mouse

Ventral view of a 14-day-old transgenic mouse, carrying a reporter gene that is active in the developing limb.

Stereomicroscopy Photo by Dr. Mohammad K. HajihosseiniUniversity of East Anglia, Norwich, U.K.Bioscapes 2009 photo contest

Page 33: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.
Page 34: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Gene therapy

Page 35: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Insertion of genes using the Ti plasmid

Page 36: Biotechnology Biology 122 Genes and Development. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction.

Genes inserted into rice to produce Golden rice


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