Name: Mckoy Sanderson

ID No. 620128253

Title: Fluctuation Test

Aim: To determine the mean and the variance value for both large and small cultures and how

they are compared according to the induced and spontaneous mutation.

Introduction:

The fluctuation test was designed to distinguish whether or not mutation was spontaneous or

induced. If the mutation is induced, the mutation must arise when the mutagen T1 is present. If

this is the case, then all the cells have equal probability of the mutation arising and so if we do

this multiple times, we are supposed to see consistent result. Hence, small fluctuation in the

results. However, if mutation is spontaneous, then mutation can exist before being exposed to the

mutagen; it can occur at any growth stage of the organisms and will do this multiple times. The

observation that will be noticed is that the results are going to differ, the results won’t be

constant, they will fluctuate. In this case, mutagen is introduced at the 4th generation. If mutation

is induced, then it will only be seen occurring at the 4th generation. As a result, the mutation rate

will remain constant, and fluctuation will be low. On the other hand, if mutation is spontaneous,

then it can occur in any stage of any generation and fluctuation will be high. During the

fluctuation test, when mutation is spontaneous, because mutation can arise at any point of the

growth phase of the bacteria, then the colonies on the plate varies. While, in the induced

hypothesis, the colonies arise only upon exposure to the bacteriophage and the cells have an

equal probability of acquiring T1 resistance upon exposure to the T1.

TABLE 1 SHOWING THE NUMBER OF T1 PHAGE RESISTANT MUTANTS IN THE

LARGE CULTURE OF E. coli CELLS FOR EACH SAMPLE.

Large Culture

Sample Number #T1R Mutants

1 13

2 12

3 10

4 17

5 17

6 15

7 20

8 19

9 11

10 21
TABLE SHOWING THE NUMBER OF T1 PHAGE RESISTANT MUTANTS FOR THE

SMALL CULTURE OF E. coli CELLS FOR EACH SAMPLE.

Small Culture

Culture #T1R Mutants

Number

1 2

2 4

3 0

4 0

5 7

6 0

7 6

8 70

9 0

10 1

11 8

12 0

13 2
14 3

15 47

16 0

17 0

18 0

19 101

20 0

TABLE 3 ILLUSTRATING WHETHER THE MUTATION WILL BE INDUCED OR

SPONTANEOUS.
 Induced Mutation Spontaneous Mutation

Variance Low High

Variance/Mean Low High

Calculations:
Discussion:

The fluctuation in number of colonies varies between the small and large cultures. The

number of T1 resistance colonies from the large culture varies slightly; the values are close (this

illustrates induced mutation). On the other hand, the T1 resistant colonies from the small culture

shows a much greater variation in the values; there were tubes that had no colonies. This

illustrates the spontaneous hypothesis, in which the number of T1 resistant cells show larger

fluctuation. The mean for the large culture was found to be 15.5, while for the large culture, it

was 12.55. these mean values are observed to be very close. However, for the small culture, the

mean values differ greatly from the T1 mutants. The variance for the large culture is close to the

mean value of the large culture, which defines the induced mutation. However, the variance for

the small culture is way higher than the variance for the large culture, and so the mean is much

less than the variance. This defines the spontaneous mutation. If the mutation is induced, it is

expected that the T1 resistant colonies arise only upon exposure to the bacteriophage and that no

matter the stage of cell growth, fluctuation should be small for both large and small cultures. If

the mutation is spontaneous, it is expected that the mutation can exist before being exposed to

the mutagen and so the results will have larger fluctuations in the number of T1 resistant cells.

The type of mutation that occurred in the large culture would be the induced mutation and the

mutation that occurred in the small culture would be spontaneous.

Conclusion:

The respective mean for the large culture and small culture was found to be 15.5 and 12.55

colonies per sample. The variance was also determined and was found to be 15.17 and 754.89 for

the large and small culture respectively. The rate of mutation was said to be spontaneous for the

small culture as it was calculated to be 8x10^-8 mutation/cell division.

References:

Burton, Z. E, & Kaguni, J. M. (1997). Experiments in molecular biology: Biochemical

application. San Diego, California.: Academic Press.

Zheng, Q. (1999). “Progress of a half century in the study of the Luria-Delbruck distribution”.

Mathematical Biosciences. 162 (1–2): 1–32. doi:10.1016/S0025-5564(99)00045-

0. PMID 10616278