1. If your PCR is inserted properly, where in the pCR 4-TOPO vector will it be inserted? 2. To use PCR to isolate the allele from the colony, you will use the primer pairs T3 and T7, or M13 F and M13 R. Look at the pCR 4-TOPO vector. Why can you use those primer pairs to isolate your alleles? 3. What do you think happend after you did the ligation and then the transformation but you don't have any bacterial colonies growing on your plate?

1. If your PCR is inserted properly, where in the pCR 4-TOPO vector will it be inserted? 2. To use PCR to isolate the allele from the colony, you will use the primer pairs T3 and T7, or M13 F and M13 R. Look at the pCR 4-TOPO vector. Why can you use those primer pairs to isolate your alleles? 3. What do you think happend after you did the ligation and then the transformation but you don't have any bacterial colonies growing on your plate?

Biology: The Dynamic Science (MindTap Course List)

4th Edition

ISBN:9781305389892

Author:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Publisher:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Chapter18: Dna Technologies: Making And Using Genetically Altered Organisms, And Other Applications

Section: Chapter Questions

Problem 1ITD: You learned in the chapter that an STR locus is a locus where alleles differ in the number of copies...

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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?
You learned in the chapter that an STR locus is a locus where alleles differ in the number of copies of a short, tandemly repeated DNA sequence. PCR is used to determine the number of alleles present, as shown by the size of the DNA fragment amplified. In the Figure below are the results of PCR analysis for STR alleles at a locus where the repeat unit length is 9 bp, and alleles are known that have 5 to 11 copies of the repeat. Given the STR alleles present in the adults, state whether each of the four juveniles could or could not be an off-spring of those two adults. Explain your answers.
1. What are denaturing DNA and how do you do it? 2. How do Primers anneal? 3. Explain what the primers are and why you need them? 4. What is the extension step? 5. How do scientist know how long the PCR product will be? - All 5 questions please!
1. Primers come at a concentration of 200 pmol//ul (uM). You want the final concentration of your primers to be 0.2 uM. How would you dilute the primers in water so that you can add 1 ul in a PCR mix with a total volume of 50ul?
Below are several problems frequently faced by researchers when running the PCR. Give one (1) solution to each problem stated. 1. No PCR product 2. Multiple bands appeared after gel electrophoresis (you are amplifying just 1 gene) 3. Bright thick bands at the end of the agarose gel after electrophoresis 4. Bands appeared at the negative control
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?
Which of the following is true for nested PCR? 1. In the second round of PCR, primers should anneal only to the target site 2. Two primers are used to ensure that one or the other will anneal. 3. Both primers anneal only to the target site, the second round is to get more DNA 4. In the first round of PCR, primers should only anneal to the target site
The temperature at which the primers and target DNA hybridize may be changed to influence the stringency of PCR amplification. What effect will changing the hybridization temperature have on the amplification? Let's say you have a certain yeast gene A and want to check whether it has a human equivalent. How might managing the hybridization's rigor benefit you?
You want to amplify a specific region of DNA during PCR. Your primers should a Both should be an exact replica of the beginning of the specific gene you want to amplify b Both should be complementary to the beginning of the specific gene you want to amplify c One primer should be identical to the beginning of the gene sequence while the other is identical to the end of the gene sequence d One primer should be complementary to the beginning of the gene sequence while the other is complementary to the end of the gene sequence e One primer should be identical to the beginning of the gene sequence while the other is complementary to the end of the gene sequence
10) Recombinant DNA technology and selective breeding are 2 techniques that seek to produce desirable traits within an organism, yet they do so in very different ways. How do the methods of each process differ? What technique wold more reliable in producing the desired outcome? Why?
1. You are studying a new gene “X” that you think controls skin color in Bearded Dragons. In order to determine what gene X does, you need lots of gene X DNA to work with. So, you decide to amplify it through the process known as PCR. You have a forward and a reverse primer that have been designed to gene Xf – a gene in frogs that is similar to the one found in the Bearded Dragon. You are pretty sure that these two primers will work in Bearded Dragons, so you decide to go ahead with the PCR. In frogs, the Forward Xf primer binds gene Xf between nucleotides 32 and 47. The Reverse Xf primer binds between nucleotides 1110 and 1135. a. What size PCR product would these two primers create if they amplified the frog gene Xf?? 3. A fellow lab worker brings you DNA containing what might be a similar gene in Leopard Geckos (XG). She asks you to see if you can amplify it using the same primers you used in frogs and the Bearded Dragon. You run the PCR and then analyze the product by…
Which of the following best describes the complete sequence of steps occurring during eve cycle of PCR? 1- The primers hybridize to the target DNA. 2-The mixture is heated to a high temperature to denature the double stranded target DNA 3- Fresh DNA polymerase is added. 4- DNA polymerase extends the primers to make a copy of the target DNA. E (2 Puan) 2, 3, 4 3, 4, 2 2, 1, 4 3, 4, 1, 2 1, 3, 2, 4