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Which of the following is a correct statement about the primers used in the ALU insertion PCR reaction?
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- You want to amplify the following sequence of DNA from a gene of interest that you are studying. To do this, you will use PCR. Design a forward and reverse primer. Show where the primers anneal to the template sequence. Template DNA: 5’ ATGACGGAATATAAGCTGGTGGTGGTGG---GGCTGCATGAGCTGCAAGTGTGTGCTCTCCTAA 3’ 3’ TACTGCCTTATATTCGACCACCACCACC---CCGACGTACTCGACGTTCACACACGAGAGGATT 5’ (PLEASE EXPLAIN STEP - BY STEP) Answer : Foward primer 5": Reverse primer 3"Ben used the following primer pairs to amplify the COI region of his DNA template: Afterwards, he used Agarose Gel Electrophoresis to visualize the PCR products. Based on the gel: a) Was he able to successfully amplify the desired region in all replicates? b) Are there errors in Ben's gel? Does he need to repeat his PCR?Ideal primers for the PCR reaction should have the following feature: they should have a high A-T content the forward primer should be palindromic to the reverse primer they should anneal rapidly only on one target in the DNA template they should have a high G-C content
- For the following short sequence of double stranded DNA, design primers (just ~ 3-4 bases) and show 2 copy cycles of PCR (refer to figure 13.25) for the amplification of this sequence of DNA (so that you have 4 double stranded DNA). 5’- GGTATTGGCTACTTACTGGCATCG- 3’ 3’- CCATAACCGATGAATGACCGTAGC- 5’You have two PCR primers: Forward- 5' TGAGCTAGGC 3' and Reverse- 5' GGTTCAGTCAG 3'. Show the binding sites of the primers to their Double strand DNA template. As the primer sizes are 10 and 11 bp, just write a 30 bp double stranded DNA (making sure the 5' and 3' ends of the double strand DNA in all 4 ends) and show where in the 30 bp double stranded DNA, these two primers would bind in correct orientation.A gel pattern displaying PCR products shows four strong bands. The four pieces of DNA have lengths that are approximately in the ratio of 1 : 2 : 3 : 4. The largest band is cut out of the gel, and PCR is repeated with the same primers. Again, a ladder of four bands is evident in the gel. What does this result reveal about the structure of the encoded protein?
- Look at each PCR component listed below. For each one, determine which steps(s) of the PCR reaction (denaturation, annealing or extension) would be directly affect if that component were missing. Taq polymerase: Oligonucleotide primers: DNA template: Deoxynucleotides (A, T, G and C): Imagine that you correctly prepare your PCR reaction mixture, but there is something wrong with the thermal cycler. Describe what would happen if: The thermal cycler was stuck on 940C: The thermal cycler cycled between 600C and 720C, but never reached 940C The thermal cycler cycled between 940C and 720C, but never reached 600You decide to use PCR to determine if you have another transposon insertion - this one on chromosome 4 and caused by a different jumping gene. Which of the PCR reagents would differ between this new reaction and the one you did for the ALU insertion PCR? Group of answer choices Primers Taq polymerase Free nucleotidesCGATTGCAGGTTATAGCG. Which of the following primer pair can be used to amplify this template with PCR? a. GCTAA and TAGCG b. GCTAA and CGCTA c. CGATT and TAGCG d: CGATT and CGCTA
- For the following short sequence of double stranded DNA and the given primers, there will be one major duplex DNA product after many cycles (imagine 10 cycles) of PCR. Provide the sequence of this one major duplex product and label the 5’ and 3’ ends of each strand. Sequence to be amplified: 5’- GGTATTGGCTACTTACTGGCATCG- 3’ 3’- CCATAACCGATGAATGACCGTAGC- 5’ Primers: 5’-TGGC-3’ and 5’-TGCC-3’You are an expert molecular biologist and you just had your regular PCR run and you are now checking your PCR products if you have amplified the correct desired DNA fragment. Upon checking the results of the electrophoresis, you noticed that you have several bands in your gel. You expected that you will only have a band with the same size as with the "X" size of the DNA ladder. Unfortunately, you have also seen bands that fall in the "W", "Y" and "Z" markers of the ladder. Assume you got a very high quality DNA template from your extraction step. A. What do you think are the possible causes of this incident? Explain briefly.You are an expert molecular biologist and you just had your regular PCR run and you are now checking your PCR products if you have amplified the correct desired DNA fragment. Upon checking the results of the electrophoresis, you noticed that you have several bands in your gel. You expected that you will only have a band with the same size as with the "X" size of the DNA ladder. Unfortunately, you have also seen bands that fall in the "W", "Y" and "Z" markers of the ladder. Assume you got a very high quality DNA template from your extraction step. B. What are the steps you will make to be able to avoid this in your next PCR run?