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Chapter 10: Problem 15

In the PCR process, if we assume that each cycle takes 5 minutes, how manyfold amplification would be accomplished in 1 hour?

Short Answer

Expert verified
There is a 4096-fold amplification after 1 hour.

Step by step solution

01

Calculate Number of Cycles

First, we need to determine the total number of PCR cycles that can be done in 1 hour. Since each cycle takes 5 minutes, and there are 60 minutes in an hour, we calculate the number of cycles by dividing the total time by the time per cycle. Number of cycles = Total time / Time per cycle = 60 minutes / 5 minutes = 12 cycles.
02

Understand Exponential Amplification

PCR amplification is exponential, meaning that with each cycle, the amount of DNA doubles. If we start with 1 initial molecule of DNA, after 1 cycle we will have 2 molecules, after 2 cycles, 4 molecules, and so on. Generally, after n cycles, the amount of DNA is equal to 2^n (where n is the number of cycles).
03

Calculate Total Amplification

Now, calculate the total amplification after 12 cycles using the exponential formula from Step 2. Substitute n with 12. Total amplification = 2^{12} = 4096. This means that after 12 cycles, the number of DNA molecules will be 4096 times the starting amount.

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Key Concepts

These are the key concepts you need to understand to accurately answer the question.

Exponential Growth
In the world of polymerase chain reaction (PCR), exponential growth plays a key role in how DNA is amplified. Exponential growth means that the quantity of DNA doubles with each cycle of the PCR process. This doubling effect leads to a rapid increase in the amount of DNA present, much faster than linear growth.
  • Each cycle produces double the amount of DNA compared to the previous cycle.
  • This is due to the fact that each new DNA strand serves as a template in the following cycles.
  • After n cycles, the number of DNA molecules can be calculated by the formula: \(2^n\).
To visualize this, consider starting with a single piece of DNA. After one cycle, you have 2. After two cycles, you have 4. After three cycles, you have 8, and so on. This rapid expansion means that in a few cycles, even small amounts of starting DNA can be amplified to much greater levels.
DNA Replication
At the heart of PCR is DNA replication, which is a natural process that cells use to copy their genetic information. During PCR, we mimic this natural process to specifically copy a chosen segment of DNA. In the laboratory setting, DNA replication is catalyzed by enzymes that build a new strand of DNA using an existing strand as a template.
  • In PCR, DNA polymerase is the main enzyme facilitating the replication process.
  • Heat is used to separate the double-stranded DNA into single strands.
  • Short DNA segments called primers are used to identify the specific segment to be copied.
  • Once the DNA is cooled, polymerase adds new nucleotides complementary to the template strands.
This cycle of heating, cooling, and duplicating continues multiple times, and each cycle leads to the exponential increase in the number of DNA molecules.
PCR Cycles
PCR cycles are the repeating segments of the PCR process and are fundamental to DNA amplification. Each cycle consists of several steps that repeatedly duplicate the DNA target section. A standard PCR cycle involves three main stages:
  • Denaturation: The DNA double helix is heated to separate the strands.
  • Annealing: The temperature is lowered to allow primers to bind to the DNA template.
  • Extension: DNA polymerase extends the primers to form a new DNA strand.
Repeating these cycles allows for the exponential increase in DNA quantity. As seen in standard laboratory practice, a PCR run can easily consist of 30 to 40 cycles, meaning the DNA can increase over a billion-fold from the original quantity within just a couple of hours. Understanding these cycles is crucial for grasping how PCR can produce such high yields of DNA in a relatively short period.

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Most popular questions from this chapter

For each of the following experimental goals, is PCR or gene cloning preferable and why? a. Isolate the same gene from 20 individuals. b. Isolate 100 genes from the same individual c. Isolate a mouse gene when you have a rat gene fragment.

A cystic-fibrosis mutation in a certain pedigree is due to a single nucleotide-pair change. This change destroys an EcoRI restriction site normally found in this position. How would you use this information in counseling members of this family about their likelihood of being carriers? State the precise experiments needed. Assume that you find that a woman in this family is a carrier, and it transpires that she is married to an unrelated man who also is a heterozygote for cystic fibrosis, but, in his case, it is a different mutation in the same gene. How would you counsel this couple about the risks of a child's having cystic fibrosis?

In Northern blotting, electrophoresis is used to resolve which biological molecules? What type of probe is used to identify the target molecule(s)?

The plant Arabidopsis thaliana was transformed by using the Ti plasmid into which a kanamycin-resistance gene had been inserted in the T-DNA region. Two kanamycinresistant colonies (A and B) were selected, and plants were regenerated from them. The plants were allowed to self-pollinate, and the results were as follows: Plant A selfed \(\rightarrow \frac{3}{4}\) progeny resistant to kanamycin \(\frac{1}{4}\) progeny sensitive to kanamycin Plant B selfed \(\rightarrow \quad \frac{15}{16}\) progeny resistant to kanamycin \(\frac{1}{16}\) progeny sensitive to kanamycin a. Draw the relevant plant chromosomes in both plants. b. Explain the two different ratios.

Why was cDNA and not genomic DNA used in the commercial cloning of the human insulin gene?
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