MBIO 2420 Lecture Notes - Lecture 1: Rebecca Skloot, Southern Blot, Eastern Blot
7 Dec 2016
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The immortal life of henrietta lacks - rebecca skloot. One of the most highly cited scientific publications. Earned sir southern a lasker award in 2005. Humans have 2 copies of gene x. Step 1 - restriction enzyme digestion: breaks it up into smaller fragments, use an restriction enzyme - ecor1. Step 2 - gel electrophoresis: separate base on size, some may have gene x some may not, now need to immobilize, transfer dna onto a substrate. Membrane: paper-like matrix, nylon or nitrocellulose, usually has a slight positive charge. Step 3 - dna denaturation: eliminate h-bonds with naoh. Step 4 - transfer dna to membrane: two methods for transferring dna to a membrane. Use buffer to transfer dna from gel to solid substance. Now need to identify dna sequence: but we need to make the dna on the substrate be permanent, bake at 80 c or use uv radiation.
