Homework Answers
DNA spread on plate = 100*0.08 ug/ul *10 ul/300 ul= 0.08/3
= 0.27 ug/ul
Transformation efficiency = transformed colonies / DNA spread on plate
= 136/0.27
Approximate transformation efficiency = 503
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During a particular transformation protocol, you used a stock of PGLO DNA with a concentration of...
Question #1. Lab 12: Transformation GFP Results Table 1. Transformation Results Color of Color of Number...
Question #1.
Lab 12: Transformation GFP Results Table 1. Transformation Results Color of Color of Number of Plate colonies under colonies under coloniesc UV light normal light co LB (-pGLO) LB/amp (-pGLO LB/amp +pGLO beige C LB/amp/ara (+pGLO) s beige Avololnose Table 2. Calculating transformation efficiency Number of transformed colonies (# transformants on LB/amp/ara plate) Transformation efficiency Mass of plasmid plated (transformants/ug plasmid) 0.156 750156 480 27 H Questions: Did you transform bacteria? Use your experimental results as evidence to...
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...
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...
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...
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...
You add (1.04x10^0) ng of plasmid DNA in 50 µL to 100 µL of competent E....
You add (1.04x10^0) ng of plasmid DNA in 50 µL to 100 µL of competent E. coli cells and incubate the mixture on ice for 20 min. Following a heat-shock treatment, the cells are incubated at 37 C for 30 min. At the end of the incubation you spread (1.2000x10^2) µL of the transformation mix on an agar plate and incubate the plate overnight at 37 C. The next morning you observe (6.8400x10^2) colonies on the plate, each colony having...
This is what we did in the experiment .First of all,1 µl of plasmid DNA was...
This is what we did in the experiment .First of all,1 µl of
plasmid DNA was added into the tube which contains competent cells
and the tube was tapped gently to mix DNA and the competent
bacteria. After that it was placed on ice for 30 minutes. Then, the
tube with the competent bacteria and plasmid were transferred to
heating block at 42 °C and the tube was leaved in there exactly 90
seconds. 0.25 ml of LB broth was...
The starting concentration of the plasmid DNA is 100 ngul and you added 2μ1 to your...
The starting concentration of the plasmid DNA is 100 ngul and you added 2μ1 to your cells. The cells were diluted to one ml prior to plating. First, determine how many transformants you would have if you plated the entire 1 ml of cell:s. Remember, you only plated half or your transformation reaction on plates 1 and 2. Add the number of transformants on plates 1 and 2, and then multiply by 2 for the total number transformants. Record that...
Cloning 2 Below is the restriction map of a 10 kb piece of DNA. Also shown...
Cloning 2
Below is the restriction map of a 10 kb piece of DNA. Also shown
below is a cloning vector which has two unique restriction enzyme
recognition sites, one for EcoRI (E) and one for HindIII (H). The
location of the kanamycin (kan) and ampicillin (amp) resistance
genes is also shown. Kanamycin and ampicillin are antibiotics that
are commonly used to select transformed E. colicells (consult the
Lab Manual for more information). Note that the HindIII site is
located...
This is what we did in the experiment .First of all,1 µl of plasmid DNA was...
This is what we did in the experiment .First of all,1 µl of
plasmid DNA was added into the tube which contains competent cells
and the tube was tapped gently to mix DNA and the competent
bacteria. After that it was placed on ice for 30 minutes. Then, the
tube with the competent bacteria and plasmid were transferred to
heating block at 42 °C and the tube was leaved in there exactly 90
seconds. 0.25 ml of LB broth was...
Question #1.
Lab 12: Transformation GFP Results Table 1. Transformation Results Color of Color of Number of Plate colonies under colonies under coloniesc UV light normal light co LB (-pGLO) LB/amp (-pGLO LB/amp +pGLO beige C LB/amp/ara (+pGLO) s beige Avololnose Table 2. Calculating transformation efficiency Number of transformed colonies (# transformants on LB/amp/ara plate) Transformation efficiency Mass of plasmid plated (transformants/ug plasmid) 0.156 750156 480 27 H Questions: Did you transform bacteria? Use your experimental results as evidence to...
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...
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...
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...
This is what we did in the experiment .First of all,1 µl of
plasmid DNA was added into the tube which contains competent cells
and the tube was tapped gently to mix DNA and the competent
bacteria. After that it was placed on ice for 30 minutes. Then, the
tube with the competent bacteria and plasmid were transferred to
heating block at 42 °C and the tube was leaved in there exactly 90
seconds. 0.25 ml of LB broth was...
The starting concentration of the plasmid DNA is 100 ngul and you added 2μ1 to your cells. The cells were diluted to one ml prior to plating. First, determine how many transformants you would have if you plated the entire 1 ml of cell:s. Remember, you only plated half or your transformation reaction on plates 1 and 2. Add the number of transformants on plates 1 and 2, and then multiply by 2 for the total number transformants. Record that...
Cloning 2
Below is the restriction map of a 10 kb piece of DNA. Also shown
below is a cloning vector which has two unique restriction enzyme
recognition sites, one for EcoRI (E) and one for HindIII (H). The
location of the kanamycin (kan) and ampicillin (amp) resistance
genes is also shown. Kanamycin and ampicillin are antibiotics that
are commonly used to select transformed E. colicells (consult the
Lab Manual for more information). Note that the HindIII site is
located...
This is what we did in the experiment .First of all,1 µl of
plasmid DNA was added into the tube which contains competent cells
and the tube was tapped gently to mix DNA and the competent
bacteria. After that it was placed on ice for 30 minutes. Then, the
tube with the competent bacteria and plasmid were transferred to
heating block at 42 °C and the tube was leaved in there exactly 90
seconds. 0.25 ml of LB broth was...
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