1. Chap. 5. Molecular tools for studying genes and gene activity Gel electrophoresis - agarose - polyacrylamide 1. size 2. purification 3. concentration Ethidium bromide; EtBr Size marker

2. Log MW

3. Fig. 5.3 0.2 Mb 2 Mb Pulsed-field gel electrophoresis

4. Fig. 5.4 SDS-polyacrylamide gel electrophoresis Roles of SDS 1.Denaturation 2.Coat negative charge Staining -DNA ; Ethidium Bromide (EtBr) -Protein ; coomassie brilliant blue Pre-stained size marker

5. Fig. 5.5

6. Fig. 5.6 Gel filtration chromatography

7. Fig. 5.7

8. Fig. 5.8

9. Fig. 5.9 densitometer

10. phosphorimaging Phosphorimaging plate vs. X-ray film - Sensitive & accurate

11. Methods of labeling DNA 1. kination; oligonucleotide H phosphatase kinase OH γ- 32 P-ATP

12. 5’-GAATTC-3’ 3’-CTTAAG-5’ G CTTAA AATTC G 2. Gap Filling GAATT CTTAA 1. DNA Polymerase 2.. α-32P-dATP, TTP

13. 4. PCR 1. Taq DNA Polymerase 2.. dNTP + α-32P-dATP A A AA A A

14. Figure 4.14 5’  3’ polymeriztion 5’  3’ exonuclease 3’  5’ exo. 3. Nick translation

15. Figure 4.13 5. Reverse transcription

16. Fig. 5.11 Non-radioactive probe

17. Capillary transfer electrophoresis hybridization autoradiography Southern blot

18. Northern blot Western blot Southwestern blot; DNA-protein interaction In situ hybridization

19. Fig. 5.17

20. Fig. 5.16 FISH In situ hybridization

21. Fig. 5.13 DNA fingerprinting Minisatellite DNA

22. Fig. 5.14

23. PCR (10 -2 ) 5 = 10 -10 Forensic science ; 법의학

24. DNA sequencing 1975, Sanger ddNTP DNAP dNTP α- 32 P-dATP

25. Figure 2.10

26. Fig. 5.19

27. By S.J.KIM

29. DNA Sequencing

30. High-throughput sequencing - Next generation sequencing - 10 Mb to 1 Gb

31. Pyrosequencing

32. Fig. 5.21 Restriction mapping

33. Fig. 5.22

34. Fig. 5.23

35. Fig. 5.24

36. In vitro mutagenesis; site-directed mutagenesis with PCR  protein engineering G m ATC bulge

37. Northern blot cf : RT-PCR

38. Reporter Gene Transcription ; lacZ, cat, luciferase

39. Luc +117 +1+1 -500-1500-1000-2000 -2175 Luc -1620 Luc -1144 -626 Luc -509 Luc -406 Luc -324 Luc -204 020000400006000080000 Relative Light Units (RLU) Luc pGL2-Basic vector Analysis of the cag-8 gene promoter activity Ji et al. BBRC 2006 luciferase

40. lacZ GFP

41. Fig. 5.36 Gel Mobility Shift Assay

42. Fig. 5.37 DNaseI footprinting

43. Fig. 5.38

44. Fig. 5.39

45. Thymidine kinase Neomycin r ES cell Knock-out mouse ; Gene targeting

46. Figure 5.39 Knock-down; RNAi = si RNA

47. Dkk KO mouse