Biochemistry 2280A Lecture Notes - Lecture 12: Southern Blot, Sanger Sequencing, Dideoxynucleotide

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To predict the function of a dna sequence by its homology to characterised genes. To identify differences between normal and mutated genes, Seeing dna base by base needs a powerful microscope or by using. The dna to be sequenced is used as a template for in vitro replication, the products of replication allow the template to be identified. In vitro only a short single stranded piece of dna is used aas a starting point. Replace helicase by temperature to denature the hydrogen bonds. Primer cannot be used as you don"t know the starting sequence. Put in a dna sequence of known bases so it can anneal to the primer. Primer is extended by dna polymerase (using free moving nucleotides) Doesn"t tell you the order of nucleotides being added cannot pause the reaction while being made. To find out modified nucleotides are used they stop the reaction when they"re being incorporated; they prevent subsequent binding.

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