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Last Class Isolation of cells Cell Fraction, Centrifuge Chromatography

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Presentation on theme: "Last Class Isolation of cells Cell Fraction, Centrifuge Chromatography"— Presentation transcript:

1 Last Class Isolation of cells Cell Fraction, Centrifuge Chromatography
Polyacrylamide Gel Electrophoresis, Western Blot Mass Spectrometry

2 Genetic Engineering

3 Key Techniques Restriction nucleases
DNA replication by vector or polymerase chain reaction Accurate Nucleic acid hybridization Gene sequencing Monitoring Gene expression

4 Restriction nucleases

5 Rejoining DNA after restriction digestion

6 Gel Electrophoresis to separate DNAs with different sizes

7 Labeling DNA in vitro (I)

8 Labeling DNA in vitro (II)

9 DNA hybridization

10 Nucleic acid hybridization for the determination of specific DNA fragment

11 Northern and Southern Blots
RNA and DNA Detection

12 Northern and Southern Blots

13 Microarray

14 DNA Cloning

15 DNA cloning and amplification

16 DNA sequencing (I)

17 DNA sequencing (II)

18 DNA sequencing (III)

19 DNA sequencing (IV)

20 DNA sequencing (V)

21 Polymerase Chain Reaction (PCR) (I)

22 Polymerase Chain Reaction (PCR) (II)

23 Polymerase Chain Reaction (PCR) (III)
Movie

24 Amplification of a gene of interest for cloning
PCR Applications Amplification of a gene of interest for cloning

25 PCR Applications Forensic Science

26 Protein Production by DNA Cloning

27 Analyzing Protein Functions With DNA Cloning Tools

28 Localization or Amplification of proteins by DNA Cloning of fusion proteins

29 Detection of protein interactions by DNA Cloning of fusion proteins (I)

30 Detection of protein interactions by DNA Cloning of fusion proteins (II)

31 Yeast two-hybrid system
Detection of protein interactions by DNA Cloning of fusion proteins (III) Yeast two-hybrid system

32 Identify Tumor Biomarkers by DNA Cloning
Phage display

33 Gene Mutation

34 Gene Mutation Digestion and Destroy

35 Visualizing Cells

36 Resolving Power

37 Light Microscope

38 Interference between light waves

39 Resolution Calculation

40 Two ways to get contrast

41 Four Types of light microscopy
Bright field, phase contrast, differential interference contrast, Dark-field microscopy

42 Fluorescence Microscope

43 Fluorescent Dyes

44 Blue: DNA; Green: microtubules; Red: centrimere
Fluorescent image Blue: DNA; Green: microtubules; Red: centrimere

45 Immunofluorescence

46 Immunostaining

47 Confocal Fluorescence Microscopy

48 The difference between conventional and confocal microscopes

49 3D reconstruction from confocal images

50 Transmission Electron microscopy (TEM, resolution 0.002 nm)

51 A root-tip cell under electromicroscopy

52 The scanning electron microscope (SEM)

53 Stereocillia from a hair cell

54 Summary of Visualizing Cells
Transmitted lights Fluorescence Electron microscopy


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