What is the size of the amplicon after 30 rounds of pcr

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Reference no: EM133476223

Question 1: Design a 12 bp forward and a 15 bp reverse primer manually (don't use online primer design tool) which can amplify the DNA fragment whose sequence is highlighted above. Write the primer sequences below. No need to check any other criteria for primer efficacy.

Question 2: What is the size of the amplicon after 30 rounds of PCR amplification using the above primers?

Question 3: How much minimum extension time in seconds is recommended to generate the amplicon using the above primers? Assume that the thermostable DNA polymerase copies at a rate of 1kb/min.

You want to generate a mutated version of the above amplicon. Using site-directed mutagenesis, you want to change the nucleotide at the 7831st position from G to an A. Write the sequence of your PCR forward and reverse primers which can be used to generate the desired fragment. Do not use an online primer design tool (may choose any length while designing the primers).

Question 5: Do you think the extension time needs to change to do the site-directed mutagenesis mentioned in part d above? Explain your reasonings.

Question 6: How to check if the base change happened after the PCR cycles?

Reference no: EM133476223

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