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lecture 4.docx - How are oligonucleotides(aka primer...

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How are oligonucleotides (aka primer) synthesized? Answer: This is a synthetic, chemical reaction called Phosphoramidite chemistry Rules: The primers always anneal at the 3’ end of the template strand The polymerase always extends from the 3’ OH, therefore it extends 5’-3’ direction In PCR the primers are DNA NOT RNA primers Primer design: Typical primers: between 18-24 nt in length, though not always the case ~30-60% C+G composition- this is very variable as it depends on the sequence you have
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