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 17QP
Summary Introduction
To describe: The reasons behind the revolutionary concept of PCR.
Introduction: A process of molecular biology which is responsible for the production of various copies of a particular segment of DNA is called “polymerase chain reaction (PCR).” Using the technique of PCR, a single segment of DNA can be amplified to produce a sufficient number of copies of that particular segment.
Summary Introduction
To describe: The two applications of PCR.
Introduction: A process of molecular biology which is responsible for the production of various copies of a particular segment of DNA is called “polymerase chain reaction (PCR).” Using the technique of PCR, a single segment of DNA can be amplified to produce a sufficient number of copies of that particular segment.
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PCR has revolutionalized biotechnology. Discuss three advantages and three disadvantages of PCR usage.
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?
Make the PCR Cocktail
This table lists the ingredients, stock reagent concentrations, and concentrations in the PCR reaction.
Prepare a "PCR cocktail" to be added to your samples to achieve these concentrations. Make enough cocktail to run nine samples.
[Four student samples + three positive controls + one negative control + one extra.]
{Hint: Remember that the concentration in the reaction is not the same as the concentration in the cocktail!]
Component
Stock Concentration
Concentration in the PCR reaction
Volume per reaction
Volume to make cocktail
Sterile water
-
-
µl
µl
PCR buffer w/ MgCl2
10x
1x
µl
µl
Nucleotide mix
10 mM
0.2 mM
µl
µl
Primer 1 (Forward)
10 µM
1.0 µM
µl
µl
Primer 2 (Reverse)
10 µM
1.0 µM
µl
µl
Taq DNA polymerase
5 U/µl
1.0 U
µl
µl
DNA template (sample)
-
~1 ng
20 µl
µl
Total
-
-
40 µl
µl
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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- (i) (ii) What is the major difference between the conventional PCR and real time PCR (qPCR) in terms of function? SyBR green and Taqman dyes are usually utilized in real time PCR. Which dye is more sensitive and accurate? Explain your answer.arrow_forwardIn PCR amplification Why is it important to know the length of the sequence you amplify?arrow_forwardPolymerase Chain Reaction (PCR) was invented by Kary Mullis in 1983. This technique had indeed facilitated research in various areas of molecular biology and genetics. You would like to amplify a particular gene fragment from the yeast genome using Polymerase Chain Reaction (PCR). What are the THREE (3) main cycles in PCR? Discuss the processes at each PCR cycle mentioned.arrow_forward
- PCR is quick, efficient and easy to perform. However, there are some situations when cell-based cloning is preferred over PCR to amplify a DNA sequence. Mention two of them.arrow_forwardTOPIC: PCR and Gene Cloning Basics Question: What are 2 possible roles of CaCl2 in the transformation process?arrow_forwardCloned 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?arrow_forward
- What's the main difference between first-generation sequencing, second-generation sequencing and whole genome amplification.arrow_forwardWhat are the different types of PCR and their product concepts? Site and explain the comparison briefly.arrow_forwardDescribe PCR. Give a real world example of when PCR may be used in the lab to solve a problem. Do a little research if nothing comes into your mind. For the toolbar, press ALT+F10 (PC) or ALT+FN+F10 (Mac). B I US Paragraph Arial x² X₂ ¶ Ⓡ ||| ||| 冈 A v 880 V Ix % $1 田田田由用arrow_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_forwardThe process of PCR essentially revolves around three phases. Briefly describe these phases and the events that occur in them. Take note the temperature on which these phases take place.arrow_forwardPCR errors during library amplification are one possible source of false positive results. If an error occurs in the first round of amplification, all the subsequent copies of that library fragment will also carry the variant, even though it is not present in the genome. However, reads from other library fragments spanning this same region will not have the variant, which means that identifying PCR copies, or clones, of library fragments during analysis can help to identify these types of errors. Based on what you know about PCR, which of the following statements would be true about the PCR clones in a sequencing library? A. PCR is subject to amplification bias, so reads derived from PCR clones will only map to regions that are not GC-rich. B. PCR is subject to amplification bias, so reads derived from PCR clones will only map to non-repetitive regions. C. PCR produces identical copies, so reads derived from PCR clones would map to the exact same location. D. PCR introduces many…arrow_forward
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