Last Update: 2 November 2017 Part - 1

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X 2 (Chi-Square) Method for Testing of Hypothesis.

The Chi-square method is used frequently in biological work specially to testing of hypothesis. This
may be used to compare the arrangement between observed and expected frequency distribution and lastly
we can decide that this hypothesis may accept or not mainly depending at 5% level of significance.
As testing of hypothesis by (X)2 chi-square mainly determine the goodness of fit, i.e. the observed
value how much fit with the expected value. Since we have agreed on a 5% level of significance. Therefore,
if such fitness occurs at least 5% to expected result, we can accept the hypothesis.
It (X)2 not only be used for samples of different sizes, but can be adapted to ratios with different
number of classes, such as 2 classes in Monohybrid cross and 4 classes in dihybrid cross. Essentially, the
(X)2 test is a mechanism by which deviations from a hypothetical ratio are reduced to a single value based on
the size of the sample this is allow to determine the probability.
A formula for (X)2 designed for a sample consisting of two classes (i.e. 1: 1 or 3 : 1 ratios) is
symbolized as follows :--
| (O1 -- E1)2| | (O2 E2)2|
X2 = [ ------------ + ------------------ - ]
E1 E2

Where O1 = Observed no. of first class.

E1 = Expected no, of first class defined from the ratio.
O2 = Observed fore 2nd class.
And E2 = Expected no. of 2nd class.

When each of these deviations is squared [(O E)2] and divided by the expected value (E) for that
class. The resulting fractions can be added () to give a single x2 value. The formula can be symbolized by
the
| (O E)2|
X = ------------
2

E
Where, (O E) is the deviation between the observed and expected class value, E is the expected
value of that respective class.

Interpretation: If the deviations of expected from the observed event are small, X2 approached to zero (0)
and the fitness is good: if the deviation is large the X2 increased and the fit is poor.

Example: Two sample, 15 : 35 and 240 : 260 on the basis of 1 : 1 of Mendelian hypothesis find out
that this may accepted or rejected.
Class - 1:
Total 15 + 35 = 50
Expected value of each class (15 + 35) 2 = 25
The deviations | (O E) | on either side of E are 10
That is, 25 15 = 10 and 25 35 = -10

Class - 2
Total 240 + 260 = 500
Expected value of each class = (240 + 260) 2 = 250
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 The deviation | (O E) | on either side of E are also here 10,
That is 240 250 = -10 and 260 250 = 10

| (O E)2| (10)2 (-10)2 200

1. X2 = ------------ = ------ + ------- = ------ = 8, df = 1
E 25 25 25

| (O E)2| (10)2 (-10)2 200

2. X = ------------ = ------- + -------
2
= ------- = 0.8, df = 1
E 250 250 250

The X2 value of 8 for smaller sample is greater then that of 0.8 for the larger sample though the
actual deviation is equal, thus, the X2 value is related to the sample size and variability within the sample.

Interpretation II: When more than two groups are classified from the sample (eg. 1: 2 : 1 or 9 : 3 : 3 : 1
ratios) each class is included in the summation, which is the X2 value.

Degrees of freedom: Generally the degrees of freedom (df) are calculated by the one less than the number
of classes.
Two class ratios (eg. 1 : 1 or 3 : 1) have one degrees of freedom and four class ratios (eg. 9 : 3 : 3 : 1)
have 3 df. and so on.
When x2 and the df. have been determined, then the x2 table may be consulted for the probability
value (p). Where it (x2 table) representing the no of df at the left and horizontally the x 2 value of each
respective df. and p value was given directly on the top line.
In our example-1, X2 of 8 for the at the df 1, the p value beyond the 0.01, i.e. > 6.635, it is indicate
that the experimental result match with the ideal/ standard/ expected result is poorly fitted i.e. fitted at 1%,
as we know that fitness less than 5% is consider as, this experimental hypothesis rejected, for this, another
hypothesis might be considered.
In the 2nd example, X2 of 0.8 at the df 1, fall between 0.455 and 1.074 or between P value of 0.50
and 0.30 this result indicated that the testing hypothesis is matched with the expected hypothesis as much as
30% - 50%, we know the testing hypothesis may be fitted with 1:1 expected ratio very well and may
accepted, and such a deviation 50% - 70% could be explain readily by chance.

For Mendelian Monohybrid Experiments:

EXAMPLE=1 TALL SHORT TOTALS
f0 30 20 50
fE x 50 = 37.5 x 50 = 12.5 50
f0-fE -7.5 7.5
(f0-fE)-1/2 -7.0 7.0
| (1f0-fE| -1/2)2 49.0 49.0
| (1f0-fE| -1/2)2/fE 49.0/ 37.5 = 1.31 49.0/ 12.5 = 3.92

X2 = 1.31 + 3.92 = 5.23

df = 2 1 = 1

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For 1:1 Medelian ratios:
EXAMPLE=1 TALL SHORT TOTALS
f0 30 20 50
fE x 50 = 25 x 50 = 25 50
f0-fE 5.0 - 5.0
| (f0-fE) | -1/2 4.5 - 4.5
| (1f0-fE1) |-1/2)2 20.25 20.25
(|f0-fE|-1/2)2/fE 20.25/ 25 = 0 .81 20.25/ 25 = 0.81

x2 = 0.81 + 0.81 = 1.62

df = 2-1 = 1

For 3:3:3:1 Mendelian ratios:

EXAMPLE=3 AB Ab aB ab Totals
f0 1080 210 200 110 1600
fE 9/16 x 1600 3/ 16 x 1600 3/ 16 x 1600 1/ 16 x 1600 1600
= 900 = 300 = 300 = 100
f0-fE 180 -90 -100 10
| (f0-fE)2| 32400 8100 10000 100
| (f0-fE)2| / fE 32400/ 900 8100/ 300 10000/ 300 100/ 100
= 36.00 = 27.00 = 33.33 = 1.00

x2 = 36.00 + 27.00 + 33.33 + 1.00 = 97.33 ; df = 4-1 =3.

For 3 df, the probability of a X2 value 97.33 or higher is fall less than 0.001 column of the X2-table. This
result is therefore relatively improbable and certainly significant at the 5% level. The null hypothesis of
observed ratio as 9:3:3:1 can therefore be rejected and thus it can be accepted.

Yets Correction:
The reduction of 1/2 or 0.5 from the absolute value to the |(O-E)| deviation is known as Yets
Correction and adds to the accuracy of X2 determination when the number of the expected classes is small.

Null hypotheses:
This hypothesis tasted that there is no differences between the observed values of the population and
if there will any difference apparently recurred it will treated as occurred by chance this difference may be
statistically proved by the testing of goodness of fit. If goodness of fit found to be statistically significant
then the hypothesis may be rejected, and when it was insignificant, then it will be accepted.

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