LEC23 - cDNA libraries can be made more selectively than...

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cDNA libraries can be made more selectively than genomic libraries . Tissue specific cDNA libraries: Eg. brain, muscle, liver, lymphocytes, kidney, testes cDNA libraries before and after differentiation Cancer cell cDNA libraries
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Genomic vs cDNA libraries
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Techniques Western blots ELISA PCR DNA sequencing
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Detection of proteins using Western blots Separate proteins by size using SDS gel electrophoresis SDS denatures protein and coats it with negative charge Smallest proteins migrate faster in gel Transfer to membrane Detect specific proteins with antibodies Detect protein-antibody using enzyme conjugated secondary antibodies 1 2 3 4
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Second antibody recognizes first antibody and is linked to either an enzyme (horse radish peroxidase (HRP)) or a fluor
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If the primary antibody was covalently linked to an HRP, would the secondary antibody be necessary? A. Yes B. No
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ELISA Enzyme-Linked Immunosorbent Assay
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Can be used to detect the amount of antibody in a blood sample
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PCR An alternative to cloning (or used in conjunction with it) PCR: Polymerase chain reaction Method to amplify specific regions of the genome in
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