Biotechnology Biology 122 Genes and Development Restriction enzymes

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 c. DNA

Colony hybridization

Southerns

RFLP analysis

PCR

Dideoxynucleotides

Manual DNA sequencing

Sequencing by synthesis Copyright © The Mc. Graw-Hill Companies, Inc. Permission required for reproduction or display. 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 Mc. Graw-Hill Companies, Inc. Permission required for reproduction or display. Bridge amplification with unlabeled d. NTPs c. Free end binds to primer Fragments become doublestranded d. Attached Free terminus

Copyright © The Mc. Graw-Hill Companies, Inc. Permission required for reproduction or display. Bridge amplification with unlabeled d. NTPs c. Free end binds to primer Fragments become doublestranded d. Attached Free terminus Denature doublestranded molecules e. Attached

Copyright © The Mc. Graw-Hill Companies, Inc. Permission required for reproduction or display. Bridge amplification with unlabeled d. NTPs 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 Mc. Graw-Hill Companies, Inc. Permission required for reproduction or display. G

Yeast Two-hybrid system

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