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- In a PCR reaction to amply a particular gene, 120 copy of the gene were initially used as DNA templates (this is the Nō). Theoretically, how many copies of this particular gene can be obtained after 25 PCR cycles? 4.03 × 10⁹ 2.01 × 10⁹ 1.02 × 10⁹ O 6.04 × 10⁹What purpose do short DNA primers serve in PCR? O They determine which sequence of DNA is copied O They synthesize new DNA O They synthesize proteins O They separate RNA from DNA MacBook Air 80 888 F7 F65' 3' O 60°C 5' O 95°C Primer 1 O 75°C ORF For your PCR reaction above, the primer set has a much higher AT (low GC) content than what would be considered normal. Due to this difference in AT content, the optimal annealing temperature for the primers is going to have to be altered from the "normal" temperature cycle: 95°C 60°C ⇒ 75°C Which of the temperatures will have to be altered to accommodate this difference the primer set? Primer 25¹ 3' 5' annealing with
- What are the reasons why there needs to be more than 10 cycles in the PCR process? Name 2 reasons.If you wanted to prepare only one PCR reaction, how much of each reagent would you add to the PCR tube?] What is the final volume of the individual PCR reactions we are making?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.
- 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.The final amount of each primer required in a PCR reaction is 25 picomol. If the total volume of the PCR reaction is equal to 100 µl and the stock concentration of each primer is equal to 0.0025 mM. Calculate the volume of stock primer that needs to be added in order to ensure a primer amount of 25 picomol.How many microliters of pGLO plasmid will you need for a PCR reaction is you need 200 nanograms and the plasmid concentration is 0.05 micrograms/microliter?
- Explain why 30–45 cycles are usually carried out in most PCR reactionsWhy does the temperature constantly fluctuate inside the PCR machine? What is the purpose of the 95oC temperature? 60oC? 72oC? (Give answers longer than a single word.)If you are trying to set up a PCR with a total volume of 40uL and you have access to a stock solution of 5x buffer, how much 5x buffer will you be adding to the PCR? ___uL?