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Gene Cloning Techniques: Academic and Corporate Strategies

Learn gene cloning methods to identify genes, such as PCR, RT-PCR, DNA and expression library screening, microarray analysis, and mutant rescue. Includes Southern and Northern blot hybridization, primer extension analysis, and PITX2-LACZ techniques.

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Gene Cloning Techniques: Academic and Corporate Strategies

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  1. How to get the cloned gene? (How to clone or identify the gene?) (1)From academic scientist (2)From company (3)PCR (RT-PCR) (4)From DNA library (Hybridization) (5)From expression library (Antibody) (6)From expression library (Protein Activity) (7) Microarray analysis (8) Mutant (Complement or Rescue)

  2. Figure 4.12. Southern hybridization. (A) Transfer of DNA from the gel to the membrane. (B) The membrane is probed with a radioactively labeled DNA molecule. On the resulting autoradiograph, one hybridizing band is seen in lane 2, and two in lane 3.

  3. Northern Blot Hybridization

  4. Primer Extension Analysis

  5. S1 Nuclease Mapping Analysis

  6. Reporter Gene Assay

  7. PITX2-LACZ Staining of mouse embryo day 11 PITX2: bicoid-related transcription factor

  8. Luciferase gene Luciferase Activity (Promega)

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