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How does PCR work? (1 Viewer)

SadCeliac

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I understand that it replicates DNA/a gene through a cyclic process (Denaturing, Annealing, Something??? --somewhere here DNA is heated and cooled?) but I don't quite know what each stage does / how it actually works. I also know that it needs lots of free nucleotides to work

Also, how would I contrast it with Recombinant Plasmid replication? E.g., for a 3 mark question
 

SadCeliac

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I understand that it replicates DNA/a gene through a cyclic process
Update: I think this summarises it, but I would like someone's clarification <3

DENATURING >> ANNEALING >> EXTENSION

PCR requirements:
- DNA sample
- primers
- free nucleotides
- Taq polymerase enzyme
- buffer

Process:
- Heat to high temp (95degrees)
- Denaturing occurs (strands separate)
- Cool to lower temp (55 degrees)
- Annealing occurs (primers bind to template DNA)
- Heat slightly (to 72degrees)
- Extension occurs (new strand is synthesised)
- Repeat
 

Big_Kev

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yep exactly - u can also apply this to Sanger sequencing (the initial steps bcoz DNA must be amplified through PCR) but yuh overall nice
 

SadCeliac

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yep exactly - u can also apply this to Sanger sequencing (the initial steps bcoz DNA must be amplified through PCR) but yuh overall nice
okok - so literally just

seperate the strands >> add primers >> extend with taq polymerase
 

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