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BIOL200 Review Techniques.docx

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Biology (Sci)
BIOL 200
Richard Roy

Lecture16- Molecular Genetics recombinant DNA technology cloning bacterial restriction enzymes Differential digestion + directional insertion Gnomic libraries genetarion cDNA libraries generation mRNA enrichment through purification cDNA generation Gnomic libraries screening cDNA libraries screening PCR RT-PCR Overexpression of recombinant protein Lipid treatment or electroporation to transfect Selection markers Fusion protein for functional analysis Fusion protein Qualitative analysis (size, nucleotide composition, conformation, structure) Quantitative analysis (level of product… ie tumor markers) Molecular probes (fluorescent, radioactive) DNA/RNA detection (nucleic acid hybridazion techniques):  Run a gel  Transferring to solid state support (alkaline solution and capillary action)  Molecular probes  Degenerate pool and polynucleotide kinase (PNK)  autoradiography Molecular identification of RFLPs Generation of radiolabelled probe (dNTPs labelled on alpha phospate) PCR in genotyping Southern Blot (DNA) Northern Blot (RNA) Western Blot (proteing) Lecture17- Eukaryotic Transcription 1 Alpha amatin resistance Lecture18- Eukaryotic Transcription 2 Linker Scanning Mutations Electrophoretic mobility shift assay (EMSA) DNA footprinting Determining modular structure of transcription factors Lecture19- RNA Processing 1
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