Homework Answers
a. Ori= origin of replication.
b. Ara pBAD= Arabinose B promoter
c. GFP= Green fluorescent protein.
d. araC= Arabinose deletion that regulates production of GFP.
e. bla ampR = Beta Lactamase ampicillin resistance. Ara C= arabinose operon activation
Add Answer to:
Question #1.
Lab 12: Transformation GFP Results Table 1. Transformation Results Color of Color of Number...
Suppose that you carried out a Bacterial transformation of E. coli HA101 with pGLO plasmid experiment...
Suppose that you carried out a Bacterial transformation of E. coli HA101 with pGLO plasmid experiment in the lab. During the experiment plates with bacteria were inoculated from +GLO and -GLO microfuge tubes (LB (-) plate, LB/amp (-) plate, LB/amp (+) plate, and LB/amp/ara (+) plate). 14) Explain what kinds of bacterial growth you will find on each of 4 plates after incubation ((LB (-) plate, LB/amp (-) plate, LB/amp (+) plate, and LB/amp/ara (+) plate): under normal light and...
.1)a) Why was it important that the –pGLO bacteria were found growing on the LB plate?...
.1)a) Why was it important that the –pGLO bacteria were found growing on the LB plate? Why would their absence create problems in interpreting our results? b) What two conclusions (one per plate) should you draw from the observation that –pGLO bacteria did not grow on the LB/amp plate, but that some +pGLO bacteria did? c). Why did we observe bacteria colonies that glowed under UV light when we inoculated +pGLO bacteria on an LB/amp/ara plate, but not when we...
1. Fill in the table above with what you observe on your plates. 2. Bacterial transformation...
1. Fill in the table above with what you observe on your
plates.
2. Bacterial transformation occurred on which agar plate (s)?
What evidence do you have that the bacteria were transformed
here?
3. Which plates have glowing growth? Explain what causes
bacteria to glow.
II. Transformation of E. coli with Plasmid DNA (PGLO) 1. Three LB (Luria Broth) agar plates are obtained. The plates contain: • Plate A: LB-Agar/Ampicillin/Arabinose • Plate B: LB-Agar/Ampicillin • Plate C: LB-Agar 2. Three...
During a particular transformation protocol, you used a stock of PGLO DNA with a concentration of...
During a particular transformation protocol, you used a stock of PGLO DNA with a concentration of 0.08 ug ul. You transformed 191 UL of competent E. coli with 100L of PGLO DNA. You added 191 UL of LB broth to the transformation before plating 100 uL of the culture to a plate labeled +POLO, LB, Amp, Ar. After growing the plates overnight, 136 fluorescent colonies (transformants) were counted. What was the average transformation efficiency of your work?
Number of colonies: Plate LB, no DNA LB/ amp, no DNA uncountable LB/ amp, DNA (The two plated lab...
Number of colonies: Plate LB, no DNA LB/ amp, no DNA uncountable LB/ amp, DNA (The two plated labeled Bacteria DNA) 683 (70 ul) 291 (3011) (a) What quantity of DNA (in ng) was used in the transformation? (2 marks) (b) What fraction of the total transformation reaction was plated out? (2 marks) (c) How many transformants (colonies) were there in the total transformation reaction? (2 marks) (d) What was the transformation efficiency, expressed as transformants per ug of DNA...
1. On which plate do you expect bacteria to glow when viewed under UV light? Which...
1. On which plate do you expect bacteria to glow when viewed under UV light? Which will grow more colonies than others? Fill in applicable options: all glow / some glow, some don't / none glow / more colonies / less colonies Plate conditions LB with Ampicillin plate LB no Ampicillin plate none glow more colonies Both gfp transformed and untransformed bacteria in normal light Both gfp transformed and untransformed bacteria in UV light
1. Which plate(s) showed an unexpected result in the above experiment? 2. Provide the most likely...
1. Which plate(s) showed an unexpected result in the above
experiment?
2. Provide the most likely explanation for the unexpected
results.
Table 2: Hypothetical results from transformation experiments performed in a pGLO plasmid added to cells? No No Type of agar plate Total # colonies # Green colonies under UV light LB LB + ampicillin LB + ampicillin LB + ampicillin+ arabinose 100 es
Results 3. (5 points) Our transformation procedure used 10 u of 0.08 ug/ul PGLO. How does...
Results 3. (5 points) Our transformation procedure used 10 u of 0.08 ug/ul PGLO. How does the concentration of DNA affect the number of colonies and the transformation efficiency? Do you think if we used more concentrated PGLO that we could have gotten a higher transformation efficiency? The numbers of colonies shown below resulted from transformation with 10ul of PGLO at the concentrations shown. a. Calculate the transformation efficiency at each concentration. Express your answers in scientific notation. Write the...
QUESTION 1 Match each item with its function in our transformation experiment. (5 points) - Makes...
QUESTION 1 Match each item with its function in our transformation experiment. (5 points) - Makes cells permeable to plasmid DNA A. CaCl2 - Selects for the growth of transformed cells B. Ampicillin - Provides the GFP gene C. PGLO plasmid • Induces GFP gene expression D. Arabinose - Causes Green Fluorescent Protein to fluoresce E. UV Light QUESTION 2 To increase the efficiency of transformation, cells will be placed in a 42°C water bath for 50 seconds during which...
Multiple Choice, Please answer all 4 What is the purpose of the neutralization solution when purifying...
Multiple Choice, Please answer all 4 What is the purpose of the neutralization solution when purifying plasmid DNA? precipitates plasmid DNA lyses the cells contains a detergent precipitates chromosomal DNA and proteins Why does the DNA bind to the purification resin? the resin is negatively charged the resin is hydrophobic the resin is positively charged the resin is neutral Why did you get colonies on plate A but not on B in the transformation, even though both plates had ampicillin?...
1. Fill in the table above with what you observe on your
plates.
2. Bacterial transformation occurred on which agar plate (s)?
What evidence do you have that the bacteria were transformed
here?
3. Which plates have glowing growth? Explain what causes
bacteria to glow.
II. Transformation of E. coli with Plasmid DNA (PGLO) 1. Three LB (Luria Broth) agar plates are obtained. The plates contain: • Plate A: LB-Agar/Ampicillin/Arabinose • Plate B: LB-Agar/Ampicillin • Plate C: LB-Agar 2. Three...
During a particular transformation protocol, you used a stock of PGLO DNA with a concentration of 0.08 ug ul. You transformed 191 UL of competent E. coli with 100L of PGLO DNA. You added 191 UL of LB broth to the transformation before plating 100 uL of the culture to a plate labeled +POLO, LB, Amp, Ar. After growing the plates overnight, 136 fluorescent colonies (transformants) were counted. What was the average transformation efficiency of your work?
Number of colonies: Plate LB, no DNA LB/ amp, no DNA uncountable LB/ amp, DNA (The two plated labeled Bacteria DNA) 683 (70 ul) 291 (3011) (a) What quantity of DNA (in ng) was used in the transformation? (2 marks) (b) What fraction of the total transformation reaction was plated out? (2 marks) (c) How many transformants (colonies) were there in the total transformation reaction? (2 marks) (d) What was the transformation efficiency, expressed as transformants per ug of DNA...
1. On which plate do you expect bacteria to glow when viewed under UV light? Which will grow more colonies than others? Fill in applicable options: all glow / some glow, some don't / none glow / more colonies / less colonies Plate conditions LB with Ampicillin plate LB no Ampicillin plate none glow more colonies Both gfp transformed and untransformed bacteria in normal light Both gfp transformed and untransformed bacteria in UV light
1. Which plate(s) showed an unexpected result in the above
experiment?
2. Provide the most likely explanation for the unexpected
results.
Table 2: Hypothetical results from transformation experiments performed in a pGLO plasmid added to cells? No No Type of agar plate Total # colonies # Green colonies under UV light LB LB + ampicillin LB + ampicillin LB + ampicillin+ arabinose 100 es
Results 3. (5 points) Our transformation procedure used 10 u of 0.08 ug/ul PGLO. How does the concentration of DNA affect the number of colonies and the transformation efficiency? Do you think if we used more concentrated PGLO that we could have gotten a higher transformation efficiency? The numbers of colonies shown below resulted from transformation with 10ul of PGLO at the concentrations shown. a. Calculate the transformation efficiency at each concentration. Express your answers in scientific notation. Write the...
QUESTION 1 Match each item with its function in our transformation experiment. (5 points) - Makes cells permeable to plasmid DNA A. CaCl2 - Selects for the growth of transformed cells B. Ampicillin - Provides the GFP gene C. PGLO plasmid • Induces GFP gene expression D. Arabinose - Causes Green Fluorescent Protein to fluoresce E. UV Light QUESTION 2 To increase the efficiency of transformation, cells will be placed in a 42°C water bath for 50 seconds during which...
Most questions answered within 3 hours.
-
16.7
At t=0s a small "upward" (positive y) pulse centered at x = 5.0
m is...
asked 8 minutes ago -
Twitter Users and News: A poll conducted in 2013 found that 52%
of U.S. adult Twitter...
asked 23 minutes ago -
How
would I know whether a given amino acid has an ionizable group or
not? please...
asked 30 minutes ago -
True or false?
True False The function of the enzyme acyl CoA
synthetase is the ATP-dependent coupling...
asked 31 minutes ago -
Nadia Corporation adjusts its debt so that its interest coverage
(EBIT/Interest) remains constant at 3. Nadia’s...
asked 33 minutes ago -
In a clinical trial, 20 out of 600 patients taking a
prescription drug complained of flulike...
asked 39 minutes ago -
7. How many types of nuclear processes can produce energy? 8.
How many types of radioactive...
asked 42 minutes ago -
For both the Sn2 and Sn1 reaction
conditions:
Structure | Rxn (Y/N) at room T° Rxn...
asked 43 minutes ago -
11. In cell N2, enter a formula using the IF function and a
structured reference to...
asked 43 minutes ago -
There is X-linked mutations in flies in this example. You need
to determine the inheritence pattern...
asked 45 minutes ago -
1) There is a 5.0 μC charge at each of 3 corners of a square
(each...
asked 56 minutes ago -
A study of 420,095 cell phone users found that
134 of them developed cancer of the...
asked 59 minutes ago