Lab Chapter 10 pGLO part two

Question 1

You should never view plates unless…

Answer 1

transilluminator shield is down

Question 2

The more cells that are transformed to produce the needed protein, the more likely

Answer 2

that the therapy will work.

Question 3

Why is the transformation efficiency calculated?

Answer 3

to help scientists determine how well the transformation is working

Question 4

The transformation efficiency gives you an indication of…

Answer 4

how effective you were in getting DNA molecules into bacterial cells

Question 5

The transformation efficiency number represents

Answer 5

the total number of bacterial cells that express the green protein divided by the amount of DNA used in the experiment

Question 6

Before you calculate the efficiency of your transformation, you will need two pieces of information:

Answer 6

1. total number of green flourescent colonies growing on LB/amp/ara plate
2. total amount of pGLO plasmid DNA in bacterial cells spread on LB/amp/ara plate

Question 7

The most direct way to determine the total number of green fluorescent cells is

Answer 7

to count the colonies on the plate

Question 8

We need two pieces of information to find out the amount of pGLO DNA in the bac-terial cells spread on the LB/amp/ara plate in this experiment:

Answer 8

(a) What was the total amount of DNA we began the experiment with, and

(b) What fraction of the DNA (in the bacteria) actually got spread onto the LB/amp/ara plates.

Question 9

the amount of pGLO DNA in the bacterial cells that were spread on the LB/amp/ara plate can be found by

Answer 9

multiply the total amount of pGLO DNA used in this experiment by the fraction of DNA you spread on the LB/amp/ara plate.

Question 10

The total amount of DNA we began with is equal to the product of the concentration and the total volume used, or

Answer 10

DNA in μg) = (concentration of DNA in μg/μl) × (volume of DNA in μl)

Question 11

each microliter of solution contained ? μg of pGLO DNA.

Answer 11

0.1

Question 12

The fraction of DNA used is calculated by

Answer 12

volume spread on LB/amp/ara plate (uL)
/
Total sample volume in test tube (uL)

Question 13

You spread ? μl of cells containing DNA from a test tube containing a total volume of ? μl of solution.

Answer 13

You spread 100 μl of cells containing DNA from a test tube containing a total volume of 510 μl of solution.

Question 14

So, how many micrograms of pGLO DNA did you spread on the LB/amp/ ara plates?

Answer 14

To answer this question, you will need to multiply the total amount of pGLO DNA used in this experiment by the fraction of pGLO DNA you spread on the LB/amp/ ara plate.

Question 15

The expected transformation efficiency for this experiment is

Answer 15

8.0 × 102 and 7.0 × 103 transformants per microgram of DNA.