BIOC 302 Lecture Notes - Lecture 19: Restriction Enzyme, Phosphodiester Bond, Polynucleotide
Document Summary
Enzymes used to manipulate dna: restricion endonucleases, enzymes that recognize speciic dna sequences (restricion sites) and then cleave the. Dna by hydrolyzing a phosphoester bond in each strand of the double helix: most restricion sites are palindromic sequences, recogniion sequence (4-6 bp in length, restrict size of dna. Staggered sicky cut cut in a diagonal across it. Sicky ends have an over hang which can sick to some other part of something (ex. another dna) Need to design oligonucleoides that will be complementary to a region of dna; primers. For a pcr reacion: you need a pair of primers: The second to the other strand in the opposite direcion. They are present in the reacion at great excess. Sample dna dntps (lots of them in there) If you want to copy a secion of dna you have 4 possible primer sites. Opposite strands and the 3" ends have to point towards each other (of the primers)
