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Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is to
- a)decrease magnesium ion concentration
- b)decrease annealing temperature
- c)use shorter primers
- d)increase extension time
Correct answer is option 'B'. Can you explain this answer?
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Most Upvoted Answer
Amplification of a DNA fragment by PCR yields only one faint band of t...
Explanation:
When performing PCR, the goal is to amplify a specific DNA fragment. However, sometimes the amplification may not be efficient, resulting in a faint band or no band at all on the agarose gel. In such cases, adjustments can be made to improve the yield of the PCR product.
Decrease Annealing Temperature:
One way to increase the yield of the PCR product is to decrease the annealing temperature. The annealing temperature is the temperature at which the primers bind to the target DNA sequence. By decreasing the annealing temperature, the primers have a better chance of binding to the target DNA, leading to more efficient amplification. A lower annealing temperature allows more mismatches between the primers and the target DNA, increasing the probability of successful binding.
Other Options:
- Decreasing the magnesium ion concentration (option A) is not likely to improve the yield of the PCR product. Magnesium ions are essential for the activity of DNA polymerase, which is responsible for DNA synthesis during PCR. Decreasing the magnesium ion concentration can impair the activity of DNA polymerase and reduce the efficiency of amplification.
- Using shorter primers (option C) may not necessarily increase the yield of the PCR product. The length of the primers should be optimized based on the target DNA sequence and the desired specificity. Shorter primers may decrease the specificity of amplification and result in non-specific products.
- Increasing the extension time (option D) may not significantly improve the yield of the PCR product. The extension time is the duration of the DNA synthesis step during PCR. Extending the duration may increase the overall amplification, but it is unlikely to have a major impact on the efficiency of amplification.
Conclusion:
In summary, the best way to increase the yield of a PCR product when only a faint band is observed on the agarose gel is to decrease the annealing temperature. This adjustment improves the binding of the primers to the target DNA and increases the efficiency of amplification.
When performing PCR, the goal is to amplify a specific DNA fragment. However, sometimes the amplification may not be efficient, resulting in a faint band or no band at all on the agarose gel. In such cases, adjustments can be made to improve the yield of the PCR product.
Decrease Annealing Temperature:
One way to increase the yield of the PCR product is to decrease the annealing temperature. The annealing temperature is the temperature at which the primers bind to the target DNA sequence. By decreasing the annealing temperature, the primers have a better chance of binding to the target DNA, leading to more efficient amplification. A lower annealing temperature allows more mismatches between the primers and the target DNA, increasing the probability of successful binding.
Other Options:
- Decreasing the magnesium ion concentration (option A) is not likely to improve the yield of the PCR product. Magnesium ions are essential for the activity of DNA polymerase, which is responsible for DNA synthesis during PCR. Decreasing the magnesium ion concentration can impair the activity of DNA polymerase and reduce the efficiency of amplification.
- Using shorter primers (option C) may not necessarily increase the yield of the PCR product. The length of the primers should be optimized based on the target DNA sequence and the desired specificity. Shorter primers may decrease the specificity of amplification and result in non-specific products.
- Increasing the extension time (option D) may not significantly improve the yield of the PCR product. The extension time is the duration of the DNA synthesis step during PCR. Extending the duration may increase the overall amplification, but it is unlikely to have a major impact on the efficiency of amplification.
Conclusion:
In summary, the best way to increase the yield of a PCR product when only a faint band is observed on the agarose gel is to decrease the annealing temperature. This adjustment improves the binding of the primers to the target DNA and increases the efficiency of amplification.
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Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is toa)decrease magnesium ion concentrationb)decrease annealing temperaturec)use shorter primersd)increase extension timeCorrect answer is option 'B'. Can you explain this answer?
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Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is toa)decrease magnesium ion concentrationb)decrease annealing temperaturec)use shorter primersd)increase extension timeCorrect answer is option 'B'. Can you explain this answer? for IIT JAM 2024 is part of IIT JAM preparation. The Question and answers have been prepared according to the IIT JAM exam syllabus. Information about Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is toa)decrease magnesium ion concentrationb)decrease annealing temperaturec)use shorter primersd)increase extension timeCorrect answer is option 'B'. Can you explain this answer? covers all topics & solutions for IIT JAM 2024 Exam. Find important definitions, questions, meanings, examples, exercises and tests below for Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is toa)decrease magnesium ion concentrationb)decrease annealing temperaturec)use shorter primersd)increase extension timeCorrect answer is option 'B'. Can you explain this answer?.
Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is toa)decrease magnesium ion concentrationb)decrease annealing temperaturec)use shorter primersd)increase extension timeCorrect answer is option 'B'. Can you explain this answer? for IIT JAM 2024 is part of IIT JAM preparation. The Question and answers have been prepared according to the IIT JAM exam syllabus. Information about Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is toa)decrease magnesium ion concentrationb)decrease annealing temperaturec)use shorter primersd)increase extension timeCorrect answer is option 'B'. Can you explain this answer? covers all topics & solutions for IIT JAM 2024 Exam. Find important definitions, questions, meanings, examples, exercises and tests below for Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is toa)decrease magnesium ion concentrationb)decrease annealing temperaturec)use shorter primersd)increase extension timeCorrect answer is option 'B'. Can you explain this answer?.
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Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is toa)decrease magnesium ion concentrationb)decrease annealing temperaturec)use shorter primersd)increase extension timeCorrect answer is option 'B'. Can you explain this answer?, a detailed solution for Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is toa)decrease magnesium ion concentrationb)decrease annealing temperaturec)use shorter primersd)increase extension timeCorrect answer is option 'B'. Can you explain this answer? has been provided alongside types of Amplification of a DNA fragment by PCR yields only one faint band of the expected size on an agarose gel. For such a sample, the best way to increase the yield of the PCR product is toa)decrease magnesium ion concentrationb)decrease annealing temperaturec)use shorter primersd)increase extension timeCorrect answer is option 'B'. Can you explain this answer? theory, EduRev gives you an
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