For the preparations with no plasmid added, do you observe a difference between the number of...
- For the preparations with no plasmid added, do you observe a difference between the number of bacteria on the plate with LB alone and the plate, which contains LB and ampicillin? Can you explain this difference?
Homework Answers
Bacteria which does not contain palsmid will not have the gene for ampicillin resistance. In this case the bacteria will not be able to grow in the presence of ampicillin.
The presence of antibiotic resistance gene imparts antibiotic resistance to bacteria so that it can grow in the presence of antibiotic.
In the plate containing media alone, the bacteria will grow because there is no antibiotic in the media. This will occur even if the bacteria is not having plasmid.
But in the plate containing media and ampicillin, the bacteria will not be able to grow because of the inhibitory effect of ampicillin. This will occur only if the bacteria is not having plasmid. If we will insert plasmid into bacteria which is having ampicillin resistant gene, then the bacteria will grow in this plate.
Because of the presence of antibiotic resistant gene in plasmid, there are differences in the number of colonies in the plate containing only nutrient media and the plate containing nutrient media and ampicillin both
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For the preparations with no plasmid added, do you
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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...
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...
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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