BIOC 302 Lecture Notes - Lecture 9: Plasmid, Reverse Transcriptase, Oligonucleotide
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. The biological role of this: to restrict the size of dna. Cttaag: makes recombinant dna (pictured right is staggered/sicky cut) Ligases join the 5"-phosphate of one dna strand to the 3"-oh of another strand. These enzymes catalyze the addiion of a new nucleoide onto the free 3" end of a polynucleoide that is annealed to a longer, complementary polynucleoide. Need to design oligonucleoides that will be complementary to a region of dna; primers. What you need for pcr: sample dna, dntp"s, dna polymerase (thermostable, primers ssdna ~18-30 nucleoides in length need a free 3" oh, mg2+ You want to map a plasmid containing a gene of interest. Ater digesing the plasmid with restricion you get the results on an agarose gel.
