[BIOL 301] - Final Exam Guide - Ultimate 32 pages long Study Guide!
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Rt-pcr or northern blot: rt-pcr: gene expression based on transcript levels in a cell, methods: (cid:1) Need special equipment/optimization for qpcr to avoid saturation when quantifying mrna: controls: (cid:1) gene specific pcr primers, without the goi (cid:1) housekeeping gene in every well to make sure you loaded each well equally. Fuse promoter with a reporter gene (gfp, gus, etc), test activity of promoter. What"s the point: design 2 oligo-dt primers that flank a choice fragment in your goi to be used in qrt-pcr. Promotor-reporter gene fusion: replace goi in the genome with a rg. Controls: (-) tissue with no reporter, (+) tissue with known reporter with known expression pattern. Pro: easier to do than in situ but like not worth it unless you want to specifically examine the promotor. In fact, just never even do promotor fusions if you can do in situ hybridization, which is always.
