Microbiology: An Introduction
12th Edition
ISBN: 9780321929150
Author: Gerard J. Tortora, Berdell R. Funke, Christine L. Case
Publisher: PEARSON
expand_more
expand_more
format_list_bulleted
Concept explainers
Question
Chapter 9, Problem 1CAE
Summary Introduction
Introduction:
Polymerase chain reaction (PCR) is an in vitro amplification technique implemented to generate many copies of a given DNA fragment.
Expert Solution & Answer
Want to see the full answer?
Check out a sample textbook solutionStudents have asked these similar questions
Pipetting
1. Explain why a micropipettor is an important instrument in biochemistry labs.
2. Describe the basic components (and function) of a micropipettor.
Bacterial Techniques
1. Define the following.
(a) Serial Dilution
(b) Streak Plating
(c) Spread Plating
Transformation
1. Define bacterial transformation and explain why it is an important method in
biochemistry labs.
2. Describe (figure, narrative) a plasmid and describe the basic components of a
plasmid.
3. What role does CaCl2 play in bacterial transformation?
4. What role does heat shock play in bacterial transformation?
Plasmid Isolation
1. Describe the roles the following play in plasmid purification.
(a) Lysis Buffer
(b) Neutralization Buffer
(c) Elution Buffer
Please help me do this step by step.
2. Restriction Enzyme Mapping - use any resources to assist you including
the hints below.
A circular plasmid molecule (12,000 total base pairs in size) was cut with a
series of restriction enzymes and the digestions were size fractionated by
agarose gel electrophoresis. Some digestions involved just one enzyme (single
digest), some combinations of two enzymes (double digests), and one utilized all
three enzymes (triple digest).
Agarose gel electrophoresis of the digestions produced bands of the following
sizes.
Enzyme
EcoRI
Hindi!!
Pstl
EcoRI and Hindill
EcoRI and Psti
Hindill and Pstl
EcoRI, Hindill, and Pst I
Bands of the Agarose Gel (size in base pairs)
2300 and 9700
4000 and 8000
12,000
800, 1500, 2500, and 7200
2300, 3200, and 6500
4000
800, 1500, 2500, 3200, and 4000
I
Draw a plasmid map showing the location of the restriction enzyme sites
relative to each other for each map. Include all 7 maps in your answer.
Chapter 9 Solutions
Microbiology: An Introduction
Ch. 9 - Compare and contrast the following terms: a. cDNA...Ch. 9 - Differentiate the following terms. Which one is...Ch. 9 - Some commonly used restriction enzymes are listed...Ch. 9 - Suppose you want multiple copies of a gene you...Ch. 9 - Which enzyme makes the smallest fragment...Ch. 9 - Describe a recombinant DNA experiment in two or...Ch. 9 - List at least two examples of the use of rDNA in...Ch. 9 - You are attempting to insert a gene for saltwater...Ch. 9 - How does RNAi silence a gene?Ch. 9 - Prob. 10R
Ch. 9 - Restriction enzymes were first discovered with the...Ch. 9 - The DNA probe, 3-GGCTTA, will hybridize with which...Ch. 9 - Which of the following is the fourth basic step to...Ch. 9 - The following enzymes are used to make cDNA. What...Ch. 9 - If you put a gene in a virus, the next step in...Ch. 9 - You have a small gene that you want replicated by...Ch. 9 - Pieces of human DNA stored in yeast cells. a....Ch. 9 - A population of cells carrying a desired plasmid....Ch. 9 - Self-replicating DNA for transmitting a gene from...Ch. 9 - A gene that hybridizes with mRNA. a. antisense b....Ch. 9 - Design an experiment using vaccinia virus to make...Ch. 9 - Why did the use of DNA polymerase from the...Ch. 9 - The following picture shows bacterial colonies...Ch. 9 - Prob. 1CAECh. 9 - Using the restriction enzyme ECORI, the following...
Knowledge Booster
Learn more about
Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biology and related others by exploring similar questions and additional content below.Similar questions
- Part A. If student counts 63 colonies on their 10^-6 dilution LB plate. What was the original concentration of their cells if they plated 100ul? Part B.If we used pGFPuv as the template for PCR positive control. This is because: a. it contains the GFP gene so it should show a product. b. It contains DNA fragments that were added to the ligation reaction. c. It is the desired plasmid we wanted to make. d. So we have a band to compare our unknown plasid to allowing us to check if the unknown is the right size.arrow_forwardhelp answer the last questionarrow_forward= Menu #1 Mol Bio Restriction An... X + Create All tools Edit Convert E-Sign Sign in Find text or tools H 3. If the PMBBS plasmid were digested with all three REs together, how many fragments would be observed, and what would be the sizes (in bp) of these fragments? Number of fragments: Sizes: 4. Draw the actual map of the PMBBS plasmid, following the style of the sample map shown in Figure 1. Make sure you show the following information: (1) relative locations of the RE sites; (2) size in base pairs of the segments of DNA between sites. (The map need not be drawn to scale.) Answer: (provide a drawing) NFL Q Search IA W ☑ 0 Al Assistant C 2 2 C 1:1 ༢ ☑ 8:58 PM 10/22/2024 Qarrow_forward
- help answer the second questionarrow_forwardDNA Extraction by Alkaline Lysis Procedure: 1. Spin 1.5 ml of cells in a microcentrifuge at maximum speed (12,000 rpm) for 20s to pellet. Remove the supernatant completely with a Pasteur pipet or a plastic pipettor tip. The spins can be performed at 4C or at room temperature. Longer spins make it difficult to resuspend cells. 2. Resuspend pellet in 100µl GTE solution and let sit 5 min at room temperature. Be sure cells are completely resuspended. 3. Add 200µl NaOH/SDS solution, mix by tapping tube with finger, and place on ice for 5 min. 4. Add 150µl potassium acetate solution and vortex at maximum speed for 2s to mix. Place on ice for 5-15 min. Be sure mixing is complete. 5. Spin 3 min at 12,000 rpm to pellet cell debris and chromosomal DNA. 6. Transfer 0.4 ml supernatant to a fresh tube, mix it with 0.8 ml of 95% ethanol or 0.4 ml isopropanol, and let sit 2 min at room temperature precipitate nucleic acids. 7. Spin at 12,000rpm for 3 min at room temperature to pellet plasmid DNA and…arrow_forwardWrite TRUE or FALSE. If false, write the word/s that make(s) the statement incorrect. 1.The primosome complex includes primase, helicase and gyrase. 2.In plasmid DNA biotechnology, plasmid inserted with a “correct" DNA sequence is transferred to bacteria for rapid.arrow_forward
- profile-image If you take a DNA photolyase- strain of E. coli, and subject it to the treatment below step 1, spread evenly on LB plate step 2, subject to UV light step 3, cover half of plate and grow in light What will be the most likely outcome if the level of UV is nearly lethal to cells on the plate? a. Only colonies on uncovered side of plate b. Only colonies on covered side of plate c. very few colonies in roughly equal numbers on both sides of platearrow_forwardDNa Mapping using Restriction enzymes lab: We will be aliquoting and delivering 5 μl of enzyme to each of the experimental tubes. What would happen if you underloaded the enzyme? i.e. you only delivered 3 or 4 μl ? What would see in your gel results?arrow_forwardDNA Extraction by Alkaline Lysis Procedure: 1. Spin 1.5 ml of cells in a microcentrifuge at maximum speed (12,000 rpm) for 20s to pellet. Remove the supernatant completely with a Pasteur pipet or a plastic pipettor tip. The spins can be performed at FC or at room temperature. Longer spins make it difficult to resuspend cells. 2 Resuspend pellet in 100pul GTE solution and let sit 5 min at room temperature. Be sure cells are completely resuspended. 3. Add 200ul NaOH/SDS solution, mix by tapping tube with finger, and place on ice for 5 min. 4. Add 150ul potassium acetate solution and vortex at maximum speed for 2s to mix. Place on ice for 5-15 min. Be sure mixing is complete. 5. Spin 3 min at 12,000 rpm to pellet cell debris and chromosomal DNA 6. Transfer 0.4 ml supernatant to a fresh tube, mix it with 0.8 ml of 95% ethanol or 0.4 ml isopropanol, and let sit 2 min at room temperature precipitate nucleic acids. 7. Spin at 12,000rpm for 3 min at room temperature to pellet plasmid DNA and…arrow_forward
- A. DNA Sequencing 1. Determine the base sequences of the sample DNA from unknown organisms by examining the bands after gel electrophoresis on photographic films. a. Unknown 1 b. Unknown 2 AGT C I I c. Unknown 3 ||| || | | AGT | || | ||| | | | || I || || ull с I AGT 2. Write down the DNA sequences of each Unknown. I I || d. Unknown 4 || |||||| AGT || ||| | || I || ||| с C ||arrow_forwardWhat's the digest map of these 4 enzymes?arrow_forwardDrag and drop the appropriate text in the gaps below. Firstly, the plasmid DNA in the E. coli is propagated through Then the bacterial cells are pelleted by centrifugation at 10,000 RPM. The media supernatant is removed and the bacterial cells are The bacterial cells are then lysed via |. Contaminating macromolecules such as protein and chromosomal DNA is removed by Overnight incubation of the bacterial culture washed in cell resuspension buffer addition of lysis buffer addition of neutralisation buffer washing of the spin column separation via agarose gel electrophoresisarrow_forward
arrow_back_ios
SEE MORE QUESTIONS
arrow_forward_ios
Recommended textbooks for you
- Biology: The Unity and Diversity of Life (MindTap...BiologyISBN:9781305073951Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa StarrPublisher:Cengage LearningHuman Heredity: Principles and Issues (MindTap Co...BiologyISBN:9781305251052Author:Michael CummingsPublisher:Cengage Learning
Biology: The Unity and Diversity of Life (MindTap...
Biology
ISBN:9781305073951
Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr
Publisher:Cengage Learning
Human Heredity: Principles and Issues (MindTap Co...
Biology
ISBN:9781305251052
Author:Michael Cummings
Publisher:Cengage Learning