Steps of PCR in sequence are?
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
Extend DNA, anneal primers, Denture DNA
Anneal primers, extend DNA, Denature DNA
Denature DNA, anneal primers, Extends DNA
Denature DNA, Extend DNA, Anneal primers
FMGE 2018