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PCR; 4. A student is using the following PCR cycle conditions to amplify a 384 bp segment...

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Chemistry

PCR;

4. A student is using the following PCR cycle conditions toamplify a 384 bp segment of DNA:

i) incubate at 95oC for 30 seconds;

ii) incubate at 53oC for 30 seconds;

iii) incubate at 72oC for 40 seconds.

(a) Describe what is happening duringeach step (i, ii, iii) of the PCR cycle.

(b) After gel electrophoresis of thePCR products, the students see three bands (212 bp, 288 bp, 866 bp)in addition to the desired 384 bp product. Which PCR step (i, ii,iii) should be modified, and how, to try and amplify only thedesired 384 bp band? Explain.

Answer & Explanation Solved by verified expert
4.4 Ratings (574 Votes)
a Describe what is happening during each step i ii iii of the PCR cycle i incubate at 95oC for 30 seconds Denaturation stepseparation of double stranded DNA to singlestranded DNA particles step Only required for DNA polymerases that require warm actuation by hotbegin PCR This progression comprises of warming the response to a temperature of 9496 C or 98 C if greatly thermostable polymerases are utilized which is held for 19 minutes Denaturation step This progression is the primary standard cycling occasion and comprises of warming the response to 9498    See Answer
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