1. Biotechnology Biology 122 Genes and Development

2. Red arrows are the cut sites; Green dots indicate the center of the sequence ←Produces blunt ends Restriction enzymes

4. Restriction analysis

8. Cloning into plasmids

12. Production of cDNA

13. Colony hybridization

14. Southerns

15. RFLP analysis

17. PCR

18. Dideoxynucleotides

19. Manual DNA sequencing

22. Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. b: © 2007, Illumina Inc. All rights reserved Adapter Adapters a.b. Flow cell 1 cm DNA DNA fragment Dense primer lawn in flow cell Sequencing by synthesis Animation of Illumina sequencing

23. Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. c.d. Bridge amplification with unlabeled dNTPs Free end binds to primer Fragments become double- stranded Free terminus Attached

24. Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Attached c.d. e. Bridge amplification with unlabeled dNTPs Free end binds to primer Fragments become double- stranded Free terminus Denature double- stranded molecules Attached

25. Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Attached Clusters c.d. e. f. 35 cycles of bridge amplification Bridge amplification with unlabeled dNTPs Free end binds to primer Fragments become double- stranded Free terminus Denature double- stranded molecules Attached

26. Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. A A T T G G C C g. First round of synthesis with labeled dNTPs

28. Yeast Two-hybrid system

29. Construction of knockout mice

31. Normal mouse HGH mouse

32. 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. Hajihosseini University of East Anglia, Norwich, U.K. Bioscapes 2009 photo contest

34. Gene therapy

35. Insertion of genes using the Ti plasmid

36. Genes inserted into rice to produce Golden rice