4. Recall that the entire genome of the bacteria was added to the PCR reaction.
ID: 252315 • Letter: 4
Question
4. Recall that the entire genome of the bacteria was added to the PCR reaction. Describe how the Taq polymerase knows where to begin replicating the DNA. 5. Once the primer anneals to the template DNA (DNA sample), how does Taq DNA polymerase "know" what nucleotides to add? 6. Your instructor will also create positive and negative control tubes for the PCR. For the negative controls, only 3 ?1 of DNAase free water (as opposed to your DNA sample) is added to each of the two PCR tubes (containing PCR mastermix and primers). Explain why this makes sense as a negative control. Would we expect to find amplified DNA fragments in this tube? If so, which ones?Explanation / Answer
Answer 4) Taq polymerase looks for the 3' hydroxyl end of the primer that is bound to a template in the genomic DNA. Meaning the binding of primer to correct sequence in the genome is must once the genome is denatured at high temperature for the PCR reaction to occur.
Answer 5) Taq polymerase identifies the nucleotide in the template sequence that is present after the 3' hydroxyl end of the primer and adds the nucleotide that is complementary to the template.
Answer 6) No, there will be no band in the neagtive control as the template is not added. This enables us to see if there is a contamination present or not. If a cand is observed, it might be due to the contamination or due to the formation of a PRIMER DIMER.
I hope you understand now. Feel free to leave a comment down below for any further query. Thank you.
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