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Biochemistry FMGE 2018 048ffb59

Steps of PCR in sequence are?

A
Extend DNA, anneal primers, Denture DNA
B
Anneal primers, extend DNA, Denature DNA
C
Denature DNA, anneal primers, Extends DNA
D
Denature DNA, Extend DNA, Anneal primers
High-Yield Explanation
PCR - method of amplifying a target sequence of DNA* Developed by Karry Mullis in 1984* Essential requirement - 1 targeted DNA 2 Two primers (synthetic oligonucleotides of 17-30 nucleotides length) that are complementary to target DNA.3 Four deoxyribonucleotide4 THERMOSTABLE DNA polymerase * Each cycle has three step (1) Denaturation - The DNA sample is first heat denatured (>90degC) to separate the two strands of the template DNA containing the target sequence(2) Renaturation or annealing -the primers, added in excess, are allowed to anneal to the DNA (typically at 50-75degC) in order to generate the required template-primer complex.(3) Synthesis / extension -he initiation of DNA synthesis occurs at 3'-hydroxyl end of each primer. The primers are extended by joining the bases complementary to DNA strands * Each cycle of PCR takes about 3-5 minutes Three stage of each cycle in relation to temperature and time

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