explain the rationales of adding chemicals that can affect DNA stability in a polymerase chain reaction (PCR). 2.Why DNA melting is required in PCR?
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1.Briefly explain the rationales of adding chemicals that can affect DNA stability in a polymerase chain reaction (PCR).
2.Why DNA melting is required in PCR? Briefly explain how PCR can be used to detect DNA mutation.
Step by step
Solved in 2 steps
- 1. Explain what primers are and what purpose they serve in a PCR reaction. Explain the main steps and temperatures of one PCR cycle. 2. Explain the purposes of the following components of gel electrophoresis: agarose, loading buffer, DNA ladder/marker, electrode buffer, electrodes. 3. Explain the expected results of the Alu sequence at PV92 that we tested, in terms of numbers of bands and their sizes. A labeled drawing would suffice.1. What happens in the denaturing step of PCR? A. What happens during the annealing step of PCR? B. What happens during the extension step of PCR? C. What temperatures is associated with each PCR step? D. Why is it necessary to have a primer on each side of the DNA segment to be amplified? (Hint: draw it out to see what happens with only one primer). E. How did the Taq DNA polymerase acquire its name? F. Why are there nucleotides (A, T, G, C) in the master mix? What are the other components of the master mix and what are their functions?1. What is the function of the DNA polymerase enzyme in the PCR? 2. What natural process is PCR based on?
- 4. Explain the importance of primers in pinpointing the region of DNA to be amplified, and discuss why primers make PCR such a powerful and useful process. 5. In PCR, what replaces the need for helicase? In PCR, what replaces the need for primase? In PCR, why doesn't the DNA polymerase denature under the high temperatures of denaturation?1. You decide to perform dideoxy sequencing on a PCR product. You add the appropriate 32P- labeled primer, DNA polymerase, DNA template (the PCR product), buffer, DNTP mix, and a small amount of one of the four ddNTPs to four reaction tubes. You run the reactions in the thermal cycler, load each reaction into a separate lane of a polyacrylamide gel, and separate the products by gel electrophoresis. In the figure below, the lanes are labeled according to the ddNTP added. ddATP ddTTP ddCTP ddGTP - Lane 1 2 3 4 a) In lane 5, draw what you would expect to see if you prepared a reaction using a nucleotide mix containing only DATP, DTTP, dCIP, and dGTP. b) In lane 6, draw what you would expect to see if you prepared a reaction using a nucleotide mix containing only ddATP, DTTP, dCIP, dGTP. ||11. Discuss if you could perform the PCR and then agarose gel electrophoresis using a coding gene instead of a VNTR.
- 14. Why denaturation stage is important in PCR? To answer the question, please explain: 3.11. Recombinant DNA Technology 189 I) what is the basic principle of PCR? 2) what are the components of PCR? 3) what are the main stages of PCR?1. Introduction: Define the following concepts and processes 1) List material used in a PCR reaction 2) List steps of PCR reaction and temperatures for each stepBriefly explain the rationales of adding chemicals which can affect DNA stability in polymerase chain reaction (PCR).
- 1. Why is each of the following reagents required to be in the PCR reaction tube? Primers, Taq DNA polymerase, nucleotide mix, reaction buffer 2. Imagine that you forgot to add nucleotides to your PCR master mix. How would this affect the outcome of your PCR reactions? a) If you made the mistake described in the previous question, which control would give you unexpected results? (In other words, if you forgot to add nucleotides, which control reaction tube would have results that are different that they would be if you had prepared them correctly?) 3. If you found that there was DNA amplification in your negative control tube, what could be an explanation for that result?1. The polymerase chain reaction (PCR) is used by scientists to amplify DNA, particularly when the quantity of DNA is very small, mixed, or contaminated with other organisms. Explain (with words) the how PCR works using a diagram to help illustrate this (show a minimum of three cycles to illustrate your point). 2. How would one go about designing primers (I am interested in the concepts you would use to accomplish this task) that would amplify a specific sequence of the "16S rDNA," that would be found in the Clostridium family? Be sure you consider the importance of both "highly conserved" and "highly variable" regions during the amplification process. 3. How many molecules (target sequence copies) will be produced by 30 PCR cycles? Assume you start with only 1 copy of the target sequence (very unlikely)? Show your work! 4. Explain how gel electrophoresis works? Why are DNA samples that are to be separated by gel electrophoresis always loaded at the cathode end of the power source? The…3. Match the following biotech tools with their applications. Some of those tools may have more than one applications. • A. Gene cloning and Recombinant DNA • B. Restriction enzyme • C. Radioactive nucleic acid probe • D. Reverse transcriptase • E. Ti plasmid. • F. Gene therapy • G. PCR • H. Gel electrophoresis • I. single nucleotide polymorphism (SNP) and Restriction fragment length polymorphism (RFLP) • J. Genomic 1. Crime scenes and paternity 2. Making copies of DNA sequence 3. GMO 4. introduce new genes into plant cells 5. alteration of an individual's DNA to treat disease. 6. Cutting DNA at target sequence 7. Detect the presence of specific gene 8. produce a DNA strand from MRNA. 9. separate DNA molecules based on size 10. methods to get the DNA fingerprint of different individual