Polymerase chain reaction (PCR) is a technique used to amplify a fragment of DNA. When conducting this procedure, all the materials necessary for DNA replication are placed in a test tule. The test tube is first subjected to extremely high temperatures, which separates the DNA strands. The separated strands can then be replicated using the molecules supplied in the test tube. Which of the following would not need to be included in the test tube for PCR? * DNA Helicase O Primer O DNA Polymerase
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- During DNA isolation, you incubated the tissue sample at 55°C for 60 minutes, followed by incubation at 95°C for 10 minutes. What happens to the DNA molecules in this last 95°C incubation? What would happen to the PCR if this step was omitted?Entire sequence below needs to beamplified by PCR and subcloned into a plasmid vector. Which of the primersequences listed underneath is the correct reverse primer (6 marks)? Copy correctsequence into your answer. Why primer e) is not the right answer (4 marks)? 5'ATCTCTATTTAATATTTATGTCTATTTAAGCCTCATATTTAAAGACAGGGAAGAGCAGAACGGAGCCCCAGGCCTCTGTGTCCTTCCCTGCATTTCTGAGTTTCATTCTCCTGCCTGTAGCAGTGAGAAAAAGCTCCTGTCCTCCCATCCCCTGGACTGGGAGGTAGATAGGTAAATACCAAGTATTTATTACTATGACTGCTCCCCAGCCCTGGCTCTGCAATGGGCACTGGGATGAGCCGCTGTGAGCCCCTGGTCCTGAGGGTCCCCACCTGGGACCCTTGAGAGTATCAGGTCTCCCACGTGGGAGACAAGAAATCCCTGTTTAATATTTAAACAGCAGTGTTCCCCATCTGGGTCCTTGCACCCCTCACTCTGGCCTCAGCCGACTGCACAGCGGCCCCTGCATCCCCTTGGCTGTGAGGCCCCTGGACAAGCAGAGGTGGCCAGAGCTGGGAGGCATGGCCCTGGGGTCCCACGAATTTGCTGGGGAATCTCGTTTTTCTTCTTAAGACTTTTGGGACATGGTTTGACTCCCGAACATCACCGACGCGTCTCCTGCTG 3'a) 5' TTCCGGAAGAAGCTTATACGG 3'b) 5' CTGTGTTCACCTAATATTCCT 3'c) 5' CAGCAGGAGACGCGTCGGTGA 3'd) 5' AGGAATATTAGTATAATCCAC 3'e) 5' GACGCGTCGGTGATGTTCGGG 3’f) 5'…Most PCR reactions do not use the more expensive types of DNA polymerase, which have DNA proofreading. How might this be a problem in accurately copying specific DNA sequences of the target gene?
- PCR (polymerase chain reaction) is an excellent method of generating copies of target DNA. If a single piece of double stranded DNA (dsDNA) is put into a PCR machine, how many dsDNA segments will there be after 3 rounds? A. 8 segments, with 2 original strands paired B. 16 segments, with 2 original strands on different segments C. 16 segments, with 2 original strands paired D. 8 segments, with 2 original strands on different segmentsChoose the correct gel electrophoretic pattern that would be seen in dideoxy sequence analysis of the DNADNA molecule shown below. pGGCGACCGATTAGTCCCATCGATGGG−OHYou Purified plasmid DNA (a 6.0 kb vector) and got the following results... DNA conc: 200 ng/µL A260/280: 1.97 A260/230: 2.15 How would you to set up a ligation reaction with 50 ng of digested DNA and a 1254 base pair PCR insert present at a concentration of 23 ng/µL. How should you pipet the necessary nucleotide components (vector and PCR insert) to obtain a 1:5 vector to insert molar ratio? Keep in mind limitations of pipets when giving your answer.(e.g. Pipetes won't go below 0.5µL)
- A student is trying to add 15.0 ng of DNA template to a 20.0 µL PCR. The DNA template is at a concentration of 65.0 ng µL-1, and the student determines that a serial dilution is required because directly adding the DNA template would require a volume less than 1.00 µL. The student wants to prepare an intermediate solution at a concentration of 15.0 ng µL-1. If the DNA template stock will be mixed with 13.0 µL of ultrapure H2O, calculate the volume (in µL) of the DNA template required to prepare the intermediate solution.For what is each of the following used in PCR: primer, DNA polymerase, 94°C?During a PCR reaction, the step in the cycle where dNTPs are added to the primer 3’ ends to fill in the strand complementary to the template by DNA polymerase is called: Annealing Extension Hybridization Melting.
- Assume the sequence below is one half of a double stranded DNA template used in a PCR reaction. The highlighted sequences indicate the region bound by primers, either on this strand or on the other complementary strand. 5' ACGTGCGACACGTATATATGTCGCGTGAGTGTAGCGTATCGCTAGAGACGCATACCTATG 3' If the sequence of the forward primer is 5' GCGACACG 3', which of the following sequences would represent the reverse primer? a. 5’ – CAGAGATCGC – 3’ b. None of these sequences would represent the reverse primer c. 5’ – GTCTCTAGCG – 3’ d. 5’ – GCGATCTCTG – 3’ e. 5’ – CGCTAGAGAC – 3’Quantitative PCR differs from regular PCR in that it uses [A] to [B] the amount of [C] in a sample. It cannot quantify [D] unless it is first made into [F]. Match each of the following to its appropriate letter: quantify, cDNA, RNA, DNA or RNA, fluorescence. 1) A 2) B 3) C 4) D 5) E Here are the choices for the questions a) quantify b) RNA c) cDNA d) fluorescence e) DNA or RNAThe PCR reaction contains deoxynucleotide triphosphates (dNTPs) in order to construct new DNA. There are four different dNTPs used in this reaction. Which answer below lists the four different dNTPs used in PCR? A) DNA polymerase, helicase, ligase, topoisomerase B) adenine, guanine, cytosine, thymine C) adenine, guanine,cytosine, uracil D) Alanine, guanine, cytosine, Theron one