Ch5 - Recombinant DNA Technology Restriction Enzymes...

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Recombinant DNA Technology Restriction Enzymes Blotting Techniques DNA sequencing Solid-phase synthesis of nucleic acids The polymerase chain reaction (PCR) Viruses subvert their host’s molecular machinery to reproduce themselves Today, we have adapted DNA plasmids from viruses to exploit bacteria and other organisms to produce substances useful to us
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Solid Phase Synthesis of Deoxyoligonucleotides 3’phosphoramidite activates nucleotide Blocking groups prevent reactions elsewhere Anhydrous conditions 10 minute cycle, ~98% coupling After: Ammonia removes blocking groups and cleaves solid support HPLC or gel separates “failed” sequences Up to ~100 nucleotides Synthetic DNA can be used as probe or primer for synthetic proteins
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Prokaryotes use Restriction Enzymes to Split Foreign  DNA: We use them to Split DNA into Specific Fragments Palindromic recognition site Some verbal palindromes: Malayalam (language of Kerala)
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Ch5 - Recombinant DNA Technology Restriction Enzymes...

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