Suppose that another transformation experiment was conducted. In this experiment

Question

Suppose that another transformation experiment was conducted. In this experiment, 5 µl of a plasmid suspension with a concentration of 0.61 ng/µl was added to 45 µl of competent cells. The mixture was incubated on ice, subjected to temperature shock and then added to 950 µl of Luria broth. The solution was then incubated for 1 hr.

An aliquot of 85 µl was spread on a plate containing ampicillin and X-gal. This plate was labeled "undiluted.”

A tube was prepared by making 70 µl of a 1/5 dilution of the transformed cells and all 70 µl of the diluted solution was spread on a second plate with ampicillin and X-gal. This plate was labeled "diluted.”

The plates were scored after incubation at 37oC for 16 hrs. Results are given below. Assume that the plasmid suspension used in this experiment was obtained from a ligation reaction in which attempts were made to insert a hippopotamus gene into the MCS of a pUC19 vector (2686 bp). (Remember that pUC19 contains an ampicillin resistance gene and the LacZ gene. The MCS is in the LacZ gene.)Round final answers to four decimal places. Do not use scientific notation.

In making the 70 µl of diluted culture, how many µl of culture was added to the tube?

Answer:

Question text

In making the 70 µl of diluted culture, how many µl of broth was added to the tube?

Answer:

Question 23

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What is the cloning efficiency (as a percentage) for the diluted plate?

Answer:

Question 24

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How much DNA (in nanograms) was used to transform the cells on the dilutedplate?

Answer:

Question 25

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What is the transformation efficiency (ug-1) for the diluted plate? (Do not use scientific notation).

Answer:

What is the transformation efficiency (ug-1) for the diluted plate? (Do not use scientific notation).

Answer:

Question 26

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What is the cloning efficiency (as a percentage) for the undiluted plate?

Answer:

Question 27

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How much DNA (in nanograms) was used to transform the cells on the undilutedplate?

Answer:

Question 28

Question text

What is the transformation efficiency (ug-1) for the undiluted plate? (Do not use scientific notation).

Answer:

Explanation / Answer

In making the 70 µl of diluted culture, how many µl of culture was added to the tube?

Answer: 3.5l

In making the 70 µl of diluted culture, how many µl of broth was added to the tube?

Answer: 66.5l

Question 23

What is the cloning efficiency (as a percentage) for the diluted plate?

Answer:20%

Question 24

How much DNA (in nanograms) was used to transform the cells on the dilutedplate?

Answer:8.54x10-3ng/ l

Question 25

What is the transformation efficiency (ug-1) for the diluted plate? (Do not use scientific notation).

Answer:11x106cfu/ g

Question 26

What is the cloning efficiency (as a percentage) for the undiluted plate?

Answer:48%

Question 27

How much DNA (in nanograms) was used to transform the cells on the undilutedplate?

Answer:0.05185ng/ l

Question 28

What is the transformation efficiency (ug-1) for the undiluted plate? (Do not use scientific notation).

Answer:29x105cfu/g

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