Biology 2581B Lecture Notes - Lecture 9: Agarose Gel Electrophoresis, Nitrocellulose, Selectable Marker

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Sticky 3" ends 3" overhang restriction enzymes have specific recognition sequences can differ in length result in specific cutting patterns fragments fit only back together if they have the same type of ends: matching sequences. Ligate blunt end to any other blunt end: but can lose a restriction site. 5" sticky ends can be ligated together if the overhang sequences match: 2 enzymes can generate same overhangs can also lose restriction site length of recognition sequences. 8 bp will cut every 48 have to make decision of length of recognition sequence you want to pick depending on cut you want. = 700, 000 fragments average length 65. 5 kb = 46, 000 fragments can also alter digestion time enzyme wont be ablet to cut at every cut site = incomplete digest complete digest = all cuts made. Fragments made by restriction enzymes separated by size in agarose gel. Dna is negatively charged and will migrate to the positive pole.

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