Human Heredity: Principles and Issues (MindTap Course List)
11th Edition
ISBN: 9781305251052
Author: Michael Cummings
Publisher: Cengage Learning
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Chapter 13, Problem 16QP
Cloned Libraries
You are running a PCR to generate copies of a fragment of the cystic fibrosis (CF) gene. Beginning with two copies at the start, how much of an amplification of this fragment will be present after six cycles in the PCR machine?
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Running a PCR
Why is a Hotstart necessary and how is it achieved? Why are the three temperatures necessary for PCR? Explain the role of each. Why does the lid of the thermocycler need to be heated? Why did we conduct an amplification reaction with no DNA? When looking at your PCR results why do you only see one band? What size were each of your PCR products?
Briefly explain the rationales of adding chemicals that can affect DNA stability in a polymerase chain reaction (PCR). Why DNA melting is required in PCR? Briefly explain how PCR can be used to detect DNA mutation.
PCR is a very useful method in biotechnology because it does not require the 6 or more different key proteins/enzymes required for DNA replication in living cells. Explain how and why this technique only requires 1 enzyme to make lots of DNA.
Chapter 13 Solutions
Human Heredity: Principles and Issues (MindTap Course List)
Ch. 13.5 - Do you think the way this issue was handled should...Ch. 13.5 - Prob. 2EGCh. 13.7 - If you were offered the chance to have the genome...Ch. 13.7 - Prob. 2EGCh. 13 - Improving the nutritional value of food has long...Ch. 13 - Improving the nutritional value of food has long...Ch. 13 - Prob. 3CSCh. 13 - What Are Clones? Cloning is a general term used...Ch. 13 - Prob. 2QPCh. 13 - Prob. 3QP
Ch. 13 - Prob. 4QPCh. 13 - Prob. 5QPCh. 13 - Prob. 6QPCh. 13 - Cloning Genes Is a Multistep Process The following...Ch. 13 - Prob. 8QPCh. 13 - Prob. 9QPCh. 13 - Cloning Genes Is a Multistep Process Which enzyme...Ch. 13 - Cloning Genes Is a Multistep Process In cloning...Ch. 13 - Prob. 12QPCh. 13 - Prob. 13QPCh. 13 - Prob. 14QPCh. 13 - Prob. 15QPCh. 13 - Cloned Libraries You are running a PCR to generate...Ch. 13 - Prob. 17QPCh. 13 - Prob. 18QPCh. 13 - Prob. 19QPCh. 13 - Analyzing Cloned Sequences A base change (A to T)...Ch. 13 - Prob. 21QPCh. 13 - Analyzing Cloned Sequences What kind of...Ch. 13 - Prob. 23QP
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- Analyzing Cloned Sequences What kind of information can a DNA sequence provide to a researcher studying a disease-causing gene?arrow_forwardWhy DNA melting is required in PCR? Briefly explain how PCR can be used to detect DNA mutation.arrow_forwardPCR & High-Throughput Sequencing A. What are the three steps of PCR, including temperatures, (or ranges?) B. What are two differences between next-gen sequencing and Sanger sequencing?arrow_forward
- Polymerase Chain Reaction (PCR) works by exposing the reaction to a series of temperature changes. In 150 words or fewer, describe 1.) the different stages of a single PCR cycle, 2.) how many cycles of PCR your reactions will undergo, and 3.) why multiple cycles are necessary.arrow_forwardIn PCr amplification If your negative control would have shown a different bp band that what was expected, what can be a possible explanation?arrow_forwardsuppose ampliTaq gold is being used to eliminate undesired amplicons in PCR. if the pH of the reaction mixture is 8.5 at 25 degree celsius and the denaturation is done at 95 degree celcius, will the PCR run be successful? why or why notarrow_forward
- 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 calculationsarrow_forwardQuantitative PCR (qPCR) has one extra ingredient to quantify DNA. Describe its structure and explain how it changes during the reaction.arrow_forwardIn pcr: How do you know if it works? If you have more than one band in a lane, why what would be the reason?arrow_forward
- 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 segmentsarrow_forwardOur PCR samples already contain loading dye, but sometimes this isn’t the case. If your samples didn’t already contain dye and you wanted to load your PCR sample onto an agarose gel, you’d need to add loading dye to the proper concentration. There is a 6X loading dye available for use; how many µl of this loading dye will you add to 10 µl of your sample so that it is at a 1X working concentration? Show your work.arrow_forwardWhy might a PCR reaction not work for a 3500bp region you want to amplify when the reaction worked for a different region of genome from the exact same extracted DNA sample (so you know the template DNA is there and the enzyme works). You ran it for 95 degrees for 5 mins followed by 35 cycles of 95 degrees- 15 seconds, 59 degrees- 30 seconds, 72 degrees- 30 seconds Then ending with a final extension- 72 degrees, 7 mins A. The extension time needs to be increased. B. The annealing temperature might need to be adjusted-- different primers have different melting temperatures. C. The forward and reverse primers should be checked to make sure their melting temperature is similar to one another (within a few degrees) and there's no errors in the sequence. D. The extension temperature needs to be adjusted. E. All of the above F. A, B and Carrow_forward
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