BIOL 5060 Lecture Notes - Lecture 15: Myc, Polyadenylation, Reading Frame
Document Summary
Distinguish between mapping and cloning: cloning gives you physical access to a physical reagent, library screen by hybridization leads to cloning, southern blot leads to knowledge, to get physical reagents . Detection- gfp, lacz, luciferase etc : gfp- microscopy to detect intracellular location. In 5" race we have knowledge of the flanking sequences so we are able to create gene-specific primers and reverse transcribe. We can isolate and amplify our gene of interest exclusively. In this, 5" race gives you effective specificity, allowing amplification of the correct flanking sequence. Race, we have no knowledge of the sequence and, therefore, are unable to create gsps, but the poly-a tail is able to act as a primer. Because we want to include the 3" unknown end of our dna product, 3". Race has to start by pairing the natural poly a tail and a complementary poly dt (and adaptor oligo) to prime it, since this end is not known.
