01:119:115 Chapter Notes - Chapter 20: Molecular Cloning, Dna Ligase, Restriction Site
28 Mar 2016
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Rna ribose but dna deoxyribose = lacks 2" oh. Dideoxyribose also lacks 3" oh group (to which next nucleoide would normally link) Dna to be sequenced denatured into single strand. In sequencing machine synthesis starts at 3" end of primer: coninues unil a dideoxyribonucleoide inserted (at random, prevents further elongaion of strand (b/c no free 3" oh) Polyacrylamide gel separates strands shorter move faster through gel luorescent detector senses color of each tag as they come through shortest to longest spectrogram: next-generaion sequencing. Fragment copied many imes by pcr (will get to later) 106 copies: each used as a template strand. Add to well a soluion of 1 of each of 4 nucleoides then washed of repeated sequenially for all 4 nucleoides. If wrong nucleoide no ppi no lash of light recorded. Patern of lashes gives sequences of fragment in each well. Patern of lashes gives sequence of fragment in each well.
