Biology 2581B Lecture Notes - Lecture 10: Wobble Base Pair, Escherichia Coli, Fibrin
14 Mar 2017
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Finding a gene sequence based on its phenotype can be done with and without mapping information. Blood clotting cascade: wound usually clotted by fibrin, people will hemophilia a are missing viii factor, fibrin not made, wound would not clot. Purified viii factor amino acid sequence is reverse translated into a coding sequence. Problem: during reverse translation there is more than one codon for a specific amino acid (cid:498)wobble position(cid:499) Make oligonucleotides: make variations of the sequence at the wobble positions of each codon to account for all different possibilities. Degenerate: a mixture of similar, but not identical sequences. Hoping that the correct nucleotide sequence will be in there. E. coli colonies containing vectors from a genomic library. Replicate onto velvet then onto a fresh plate. Make primers to vector backbone, just on the outside of the insert. In different samples attach primer and sequence in different directions.
