MCB 2050 Study Guide - Comprehensive Final Exam Guide - Protein, Gene, Cell Surface Receptor
20 Nov 2018
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Reporter plasmid: mcs allows insertion of potential gene regulatory sequences, reporter gene codes for a protein that can be visualized or measured, luc+ = firefly luciferase gene. Basic techniques used to clone genes: construction and screening of dna libraries. Summary of major steps: digestion of vector, digest genomic dna, annealing and ligation, transformation of e coli, selection. Construction of cdna libraries: cdna is synthesized from mrna using a primer (usually oligo-dt primer since mrnas all contain a 3" polya tail) and the rna dependent dna polymerase reverse transcriptase, (synthesizes dna from an. Techniques for dna/rna analysis: gel electrophoresis: electrophoresis: uses an electric field to separate macromolecules in agarose (dna/rna) or polyacrylamide (dna or protein) gels. Dna and rna electrophoresis: since dna is negatively charged, it will migrate toward the anode (positive electrode, their rates of migration are inversely proportional to their sizes (i. e. lengths- shorter ones move faster than the longer ones.
