BIOC 2580 Study Guide - Final Guide: Exonuclease, Ecori, Base Pair
16 Dec 2015
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Type iip: symmetric recognition site and cleavage is within the recognition site. Recognizes the sequence and cuts within that sequence. If ligation is done while the restriction enzyme is still present, it will just re-cut the same site and it will be a futile cycle. Uses type iis restriction enzymes ability to cleave outside the recognition sequence to remove the sequence from the cleaved fragment. This allows cleavage and ligation to occur simultaneously. Fragment specific overhangs allows orderly assembly of multiple. Fragment specific overhangs allows orderly assembly of multiple fragments. Needs a 5" to 3" exonuclease, dna polymerase and ligase. Exonuclease chews back 5" ends resulting in complementary fragment ends to anneal and gap is filled in. Can be performed in a single tube with a master mix of enzymes and all components of reaction. Know that synthetic biology is much, much better than the old technique to quickly develop the vaccine.
