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

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

Name: Pipetting1. Explain why a micropipettor is an important instrument in
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Description: Pipetting1. Explain why a micropipettor is an important instrument in biochemistry labs.2. Describe the basic components (and function) of a micropipettor.Bacterial Techniques1. Define the following.(a) Serial Dilution(b) Streak Plating(c) Spread Pl

Biology: The Unity and Diversity of Life (MindTap Course List)

14th Edition

ISBN:9781305073951

Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Chapter8: Dna Structure And Function

Section: Chapter Questions

Problem 3DAA: HersheyChase Experiments The graph shown in FIGURE 8.5 is reproduced from an original 1952...

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biotechnology lab class :1. The bacterial streaking on antibiotic containing agar plates has been completed and the plates will be available for you to inspect in the lab class.2. You will complete a restriction enzyme digest on the four available plasmid stocks and then examine the products by agarose gel electrophoresis. Photographs of the gel are attached to identify the plasmid present in each analysed sample.Now consider the following questions :- For each of the four plasmids used in the practical, identify the size (in kb) of the linear DNA fragment(s) that you would expect to obtain if the EcoRI digest is complete.- Using the provided negative image of the gel, which shows the bands present in the ND and D samples for each plasmid, identify the plasmid that was present in each of the four plasmid stock.- Explain how you were able to identify plasmid in each sample using the gel, considering how linear DNA fragments of different length are separated by agarose gel…
In bacterial transformation ... 1, Why do you have to incubate the competent bacterial cells on ice after the addition of plasmid DNA? 2. Why do you have to add SOC (Super Optimal Broth with Catabolite repression) medium to bacterial cells? 3. Why do you have to incubate the bacterial cells in SOC medium at 37oC for 45 minutes?
Kpnl 4. Plasmid Z has a size of 7 kb, and the map shows Kpnl (K) and Pstl (P) cut sites relative to each other. This plasmid was digested with three different restriction enzymes 2000 bp 3500 bp Kpnl (K), Pstl (P) and Bgll (B) either alone or in combination and the samples run on an agarose gel as shown below. Where does Bgll (B) cut this plasmid ? Does the plasmid have one recognition site or two for Bg|l? Describe the Bgll cut site in this plasmid relative to the Kpnl cut Plasmid Z -7 kb Pstl site. How many bases to the left or right of the Kpnl cut site would you observe the Bgll cut site. Explain briefly. 1500 bp Pstl Ladder Kpni Psti K/P Bgl K/B KPB 7000 bp 7000 bp 5600 bp 5500 bp 4900 bp 3500 bp 2000 bp 1500 bp 1500 bp 1500 bp 1400 bp 600 bp %3D
ation plasmids? 9. Predict and explain the effect of each of the following conditions on the Tm of native DNA. Will the Tm be raised or lowered relative to the DNA in Tris-HCl buffer, pH 7.5? (a) Measure Tm in pH 12 buffer. (b) Measure Tm in distilled water. (c) Measure Tm in 50% methanol water. (d) Measure Tm in standard Tris-HCI, pH 7.5 buffer solution containing SDS.
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.
PCHEM4321. An agarose gel electrophoresis pattern of the plasmid PSPM4321 digestion (restriction) is shown below. Draw a restriction map of a plasmid with the appropriate restriction sites based on the data given below. Hindlll Hindll BamHI +BamHI Figure 1: 1% agarose gel electrophoresis of pCHEM4321 40 24 16 12 12 8 4 4 + |
Arial BIUA 11 + .. | I 1 I 3 I 4 i.) Fill in the table for each of the E. coli: (0) = No Activity (+) = Basal Activity and (+++) = High Activity E. coli chromosome F' Plasmid B-gal activity? Permease activity? When When When When Glucose is Lactose is Glucose is Lactose is present present present present a.) I+ P+ O+ Z+ Y+ Inone +++ +++ b.) I^[S] P+ O+ Z+ Y+ none c.) I+ P+ O^[c] Z+ Y+ none d.) I+ P+ O- Z- Y+ none e.) I+ P+ O+ Z+ Y+ I^[S] P+ O+ Z+ Y+ f.) I^[S] P+ O+ Z- Y+ I+ P+ O^[c] Z+ Y- g.) I^[TB] P+ O+ Z+ Y 1+ P+ O^[c] Z- Y+ h.) I+ P+ O^[c] Z+ Y- I+ P+ O+ Z** Y+ i.) I^[TB] P+ O^[c] Z+ Y- 1+ P+ O+ Z- Y+ Z** is a polar mutation ii. ) If the lac operon in 'a' carried a mutation in the CAP binding site that rendered it nonfunctional, how would that affect the level of ß-galactosidase protein activity with and without lactose present, why? MacBook Air 000
Complete the following tasks. You discovered that a species of bacteria can break down StyrofoamT (polystyrene) products due to an enzyme it produces, polystyrenase. You wish to study the gene that codes for this enzyme. Task 1: DNA Extraction To begin work on the bacterium, you begin by extracting its genomic DNA (GDNA). What is the purpose of the following procedures? Answer briefly but completely. Using sodium dodecyl sulfate, a detergent Answer: а. b. Adding RNase A and Proteinase K during extraction Answer: c. Adding ethanol before recovering the DNA extract С. Answer: Task 2: Polymerase Chain Reaction After purifying the gDNA extract, you want to isolate and amplify the polystyrenase gene. You perform PCR using the appropriate gene-targeted primers. What is the purpose of the following PCR components? Answer briefly but completely. DNA polymerase isolated from Thermus aquaticus Answer: а. b. Deoxynucleotide triphosphates (DNTPS) Answer: С. Forward and reverse primers Answer: Task…
AGAROSE GEL ELECTROPHORESIS PRELAB ANSWER ALL QUESTIONS 1. Type of nucleotide is one of the factors that influence electrophoretic mobility. a) True b) False 2. Electrophoresis is used for DNA separation and not proteins. a) True b) False 3. DNA Polymerase is an enzyme to cut DNA into fragments for electrophoresis. a)True b)False 4. Electrophoresis apparatus consists of Gel buffer, chamber and DNA separator. a) True b) False 5. During electrophoresis, DNA fragments collect at the Cathode. a) True b) False
A.How could endonucleases interfere with the transformation procedure? B. Does supercoiled or nicked plasmid get transformed more efficiently? Why?
2. pBR3322, an artificially made plasmid, is shown below. ECORI Hindill BamHI Pstl Amp ORI ↓↓ Tetr pBR322 (4.36 kb) ↑ Pvull Xmalll Describe 3 features that this plasmid has and indicate each one on the diagram. i) ii) 111]
Bacterial Identification Virtual Lab Add the Master Mix and answer the following questions: 13. What does the Master Mix contain? 14. What are primers? Why is a primer added? 15. Once the primers bind, what occurs next? 16. What does "highly conserved" mean? 17. Why are highly conserved regions important in this lab?