What do you think will happen if the melting temperatures of the PCR primers (forward and reverse) are far apart?
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What do you think will happen if the melting temperatures of the PCR primers (forward and reverse) are far apart?
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- What are the reasons why there needs to be more than 10 cycles in the PCR process? Name 2 reasons.You are working in a molecular biology laboratory and are having challenges with your PCR. You decide to double-check the PCR protocol programmed into the thermal cycler and discover that the annealing temperature was programmed to be 65 °C instead of 50 °C, as you had intended. What effects would this mistake have on the PCR reaction? Suggest potential solution(s).What is the difference between PCR and real-time PCR, and how do you explain it?
- What is expected theoretical number of copies of DNA molecules after 28 cycles in a PCR experiment? What is the percent efficiency of the PCR experiment if the actual number of copies was 500,000,000? Show your calculationsWhat is real-time PCR?In a typical PCR reaction, describe what is happening in stages occurring at temperature ranges (a) 92–95°C, (b) 45–65°C, and (c) 65–75°C.
- In a typical PCR reaction what is happening in stages occurring at temperature ranges (a) 92–95°C, (b) 45–65°C, and (c) 65–75°C.You want to set up 50 µl total volume of a PCR reaction. You have a microfuge tube of Taq polymerase (5 units/ul), and each PCR reaction requires a final of 10 units of Taq polymerase. You have a microfuge tube of Taq Buffer (5X), and each PCR reaction requires a final of 1X Taq Buffer. How much of Taq polymerase and Taq buffer would you add? Select all that apply 10 ul of Taq buffer 2 ul of Taq polymerase 5 ul of Taq buffer 5 ul of Taq polymeraseYou want to set up 50 µl total volume of a PCR reaction. You have a microfuge tube of Forward primer (25 µM), and each PCR reaction requires a final of 1 uM of primer. You have a microfuge tube of Reverse primer (25 µM), and each PCR reaction requires a final of 1 µM of primer. How much of the primers would you add? Select all that apply 2 ul of Reverse primer 2 ul of Forward primer 5 ul of Forward primer 5 ul of Reverse primer
- You are performing PCR for the first time using some new primers that are 25 nucleotides in length with an estimated melting temperature of 65 0C to amplify a 200 nucleotide gene. After PCR, you run an agarose gel on the samples and observe a faint band at 25 nucleotides. What is the best possible explanation for the results? The primers hybridized to multiple sites on the DNA template. Too much template DNA was added to the PCR mixture. The primers dimerized preventing DNA transcription from occurring. A nuclease was in the solution causing degradation of the DNA.what does it mean if our Pcr product was 6.1 nanograms per micrometers and our pcr script was 4.9 nanogramse per micrometers. we are doing a cloning experiment with Ldh dnaAn analyst designed a pair of primers to detect the presence of Alu sequence in human DNA genome extract. The forward and reverse primers have the melting temperature at 70 and 50 Celsius degree respectively. Will the PCR work with the following PCR condition? Denaturing: 95 C for 1 min Annealing: 55 C for 1 min Extension: 72 C for 2 min Cycling: 35 cycles Yes. Annealing temperature at 55 is a standard in PCR. Yes. Difference in meting temperature of primer pair does not affect PCR. O No. There is chance that one of the primer won't anneal to the correct sequence. O No. DNA polymerase does not work at 72 Celsius degree.