Design of Experiments:

1)Experiment: An operation which can produce some well defined results is known as
experiment.
Through experimentation, we study the effect of changes in one variable (such as
application of fertilizer) on another variable (such as grain yield of a crop).The variable
whose changed we wish to study may be termed as a dependent variable or response
variable (yield).The variable whose effect on the response variable are termed as an
independent variable or a factor. Thus, crop yield, mortality of pests etc. are known as
responses and the fertilizer, spacing, irrigation schedule, pesticide etc. are known as
factors.
2)Design of Experiments: Choice of treatments, method of assigning treatments to
experimental units and arrangement of experimental units in different patterns are known as
design of experiment.
3)Treatment: Objects of comparison in an experiment are defined as treatments. Or
Any specific experimental conditions/materials applied to the experimental units are
termed as treatments.
Ex: Different varieties tried in a trail, different chemicals, dates of sowing, and
concentration of insecticides.
A treatment is usually a combination of specific values called levels.
4)Experimental material is the objects or group of individual or animal etc… on which we
the experiment is conducted is called as experimental material. Ex: Land, Animals, lab
culture, machines etc…
5)Experimental unit: The ultimate basic object to which treatments are applied or on
which the experiment is conducted is known as experimental unit.
Ex: Piece of land, an animal, plots, etc...
6)Experimental error is the random variation present in all experimental results.
Response from all experimental units may be different to the same treatment even under
similar conditions, and it is often true that applying the same treatment over and over again
to the same unit will result in different responses in different trials. Experimental error does
not refer to conducting the wrong experiment. These variations in responses may be due to
various reasons such as factors like heterogeneity of soil, climatic factors and genetic
differences, etc.. also may cause variations (known as extraneous factors). The unknown
variations in response caused by extraneous factors are known as experimental error.
For proper interpretation of experimental results, we should have accurate
estimate of the experimental error. If the experiment errors are small we will get the
more information from an experiment, we say that the precision of the experiment is
more.
Our aim of designing an experiment will be to minimize this experimental error.
7)Layout: The placement of the treatments on the experimental units along with the
arrangement of experimental units is known as the layout of an experiment.
 Analysis of Variance (ANOVA):

The analysis of variance is a powerful statistical tool for tests of significance of

several populations mean. The term Analysis of Variance was introduced by Prof. R.A.
Fisher to deal with problems in agricultural research.
The test of significance based on Z-test and t-test are only an adequate procedure
for testing the significance of one or two sample means. In some situation, three or more
population mean to be consider at a time for testing. Therefore, an alternative procedure
is needed for testing these means. For ex: five fertilizers are applied to four plots of
wheat and its yield on each of the plot is given. We may be interested in finding out
whether the effect of these fertilizers on the yields is significantly different i.e. all the
fertilizers application on wheat plot gives same yield or different yield. Answer of this
problem is provided by the technique of analysis of variance. Thus basic purpose of
the analysis of variance is to test the equality of several means.
Variation is inherent in nature. The total variation in any set of numerical data is
due to a number of causes which may be classified as: (i) Assignable causes and (ii)
Chance causes. The variation due to assignable causes can be detected and measured,
whereas the variation due to chance causes is beyond the control of human hand and
cannot be traced separately.

Definition of ANOVA:
The analysis of variance is the systematic algebraic procedure of decomposing (i.e.
partitioning) overall variation ( i.e. total variation) in the responses observed in an
experiment into different component of variations such as treatment variation and error
variation. Each component is attributed identifiable cause or source of variation.

Assumptions of ANOVA:
For the validity of the F-test in ANOVA the following assumptions are made.
1. The effects of different factors (treatments and environmental effects) are additive in
nature.
2. The observations and experimental errors are independent
3. Experimental errors are distributed independently and normally with mean zero and
constant variation i.e. ~K(0, σ2)
4. Observations of character under study follow normal distribution
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STA541B – Design of Experiments

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Essential Reading:

Montgomery D.C, Design and Analysis of Experiments, John Wiley and Sons Inc,.
New York, 2014.

Gupta S.C and Kapoor V.K, Fundamentals of Applied Statistics, 4th edition (Reprint),
Sultan Chand and Sons, India, 2019.

K.A. Gomez and A.A. Gomez, Statistical Procedures for Agricultural Research,
Second Edition. John Wiley & sons.

Recommended Reading:

Mukhopadhyay P, Mathematical Statistics, 2nd edition revised reprint, Books and

Allied(P) Ltd, 2015.

Lawson J, Design and Analysis of Experiments with R, CRC Press, 2015.

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Definition:
Design of Experiment

Systematic procedure of planning for experiments so that the data

obtained can be analysed to yield valid and objective conclusions.

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Design of Experiments (DOE) mathematical methodology used for

planning and conducting experiments as well as analysing and
interpreting data obtained from the experiments.

It is a branch of applied statistics that is used for conducting scientific

studies of a system, process or product in which input variables (Xs)
were manipulated to investigate its effects on measured response
variable (Ys).

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Over past two decades, DOE was a very useful tool traditionally used
for improvement of product quality and reliability.

The usage of DOE has been expanded across many industries as part of
decision making process either along a new product development,
manufacturing process and improvement. It is not used only in
engineering areas it has been used in administration, marketing,
hospitals, pharmaceutical, food industry, energy and architecture, and
chromatography.

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Basic Terminologies:
Experiment: An operation which can produce some well defined results
is known as experiment. Where such inquiry will aid in on
administrative decision such as recommending or not recommending,
etc…

Through experimentation, we study the effect of changes in one variable

(such as application of fertilizer) on another variable (such as grain yield
of a crop).The variable whose changed we wish to study may be termed
as a dependent variable or response variable (yield).The variable
whose effect on the response variable are termed as an independent
variable or a factor. Thus, crop yield, mortality of pests etc. are known
as responses and the fertilizer, spacing, irrigation schedule, pesticide etc.
are known as factors

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Types of Experiments
1. Preliminary Experiment
2. Critical Experiment
3.Demonstrational Experiment

In a preliminary experiment, the investigator tries out large number of

treatments in order to obtain leads for the future work. Here most
treatments appears once

In a critical experiment, the investigator compares responses to

different treatments using sufficient observations of the responses.

Demonstrational experiments are performed when an investigator

compare a new treatment or treatments with a standard

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2) Design of Experiments: Choice of treatments, method of assigning

treatments to experimental units and arrangement of experimental
units in different patterns are known as design of experiment.

3) Treatment: Objects of comparison in an experiment are defined as

treatments. Or
Any specific experimental conditions/materials applied to the
experimental units are termed as treatments.

Ex: Different varieties tried in a trail, different chemicals, dates of

sowing, and concentration of insecticides.

A treatment is usually a combination of specific values called levels.

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4) Experimental material is the objects or group of individual or

animal etc… on which the experiment is conducted is called as
experimental material.
Ex: Land, Animals, lab culture, machines etc…

5) Experimental unit: The ultimate basic object to which treatments

are applied or on which the experiment is conducted is known as
experimental unit.
Ex: Piece of land, an animal, plots, etc...

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6) Experimental error is the random variation present in all

experimental results. Response from all experimental units may be
different to the same treatment even under similar conditions, and it is
often true that applying the same treatment over and over again to the
same unit will result in different responses in different trials.
Experimental error does not refer to conducting the wrong experiment.
These variations in responses may be due to various reasons such as
factors like heterogeneity of soil, climatic factors and genetic
differences, etc.. also may cause variations (known as extraneous
factors). The unknown variations in response caused by extraneous
factors are known as experimental error

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Case Study:
Researcher wants to study the durability (Tensile Strength)
of fabric under repeated washing using water with different
temperature (Hot, warm and Cold) and cleansing agent
(Regular, low phosphate, liquid).

● Responses variable ?
● Treatment or factor ?
● Experimental unit ?
● Level ?

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Historical perspective:

One Factor At a Time (OFAT) was very popular scientific method dominated until
early nineteen century. In this method one variable/factor is tested at a time.

In the 1920s and 1930s Ronald A. Fisher conducted a research in agriculture with
the aim of increasing yield of crop in the UK. He came up with design of experiment
and officially he was the first one who started using DOE.

In 1935, he wrote a book on DOE, in which he explained how valid conclusion

could be drawn from the experiment in presence of nuisance factors. He analysed
presence of nuisance factors with fluctuation of weather conditions (temperature,
rainfall, soil condition).

Credit for Response Surface Method (RSM) belongs to George Box who is also
from the UK. He was concerned with experimental design procedures for process
optimization.

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In 1550s, W. Edwards Deming was concerned with design of

experiment as well as statistical methods.

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Main uses of DOE:

Design of experiment is multipurpose tool that can be used in various situations for
identification of important input factors (input variable) and how they are related to
the outputs (response variable).

DOE is basically regression Model that can be used in various situations.

1. Comparison - multiple comparisons to select the best option that uses t‒test,
Z‒test, or F‒test.
2.Variable screening ‒ Intended to select important factors (variables) among many
that affect performances.
3.Transfer function identification ‒ Identified, the relationship between the input
variables and output variable can be used for further performance exploration via
transfer function.
4.System Optimization ‒ optimization by moving the experiment to optimum setting
of the variables.

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General practice and guideline for planning and conducting of DOE

1. State the objective under study

2. Define response variable under study

3. Determine the factor and the levels

4. Determine the Experimental Design

5.Perform Experiment

6. Data Analysis

7. Practical conclusion and Recommendation

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Analysis of Variance (ANOVA)

Introduction:
The analysis of variance is a powerful statistical tool for tests of significance of
several populations mean. The term Analysis of Variance was introduced by
Prof. R.A. Fisher to deal with problems in agricultural research.

Basic purpose of the analysis of variance is to test the equality of several

means.
Variation is inherent in nature. The total variation in any set of numerical data
is due to a number of causes which may be classified as:
(i) Assignable causes and (ii) Chance causes. The variation due to assignable
causes can be detected and measured, whereas the variation due to chance
causes is beyond the control of human hand and cannot be traced separately

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Definition of ANOVA:
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One-way Classification: (One-way ANOVA)
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Model (One way ANOVA):

●

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Classes / Total Mean

Groups
1 ……..

2 ……..

3 ……..

. . . . ….
. . . . …..
k ……..

Grand
Total
(GT)

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Test Procedure: The steps involved in carrying out the analysis are:

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ANOVA for CRD

Source of Variation DF Sum of Mean F ratio

Squares Squares
Treatments k-1 SSTr

Error n-k ESS

Total n-1 TSS

4) Critical value of F or Table value of F:

The table value is obtained from F-table for (k-1, n-k) df at α % &
denoted it as F tab

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● A manufacturing company has purchased three new machines of

different makes and wishes to determine whether one of them is faster
than the others in producing a certain output. Five hourly production
figures are observed at random from each machine and the results are
given below.
Machine A1 Machine A2 Machine A3
25 31 24
30 39 30
36 38 28
Observations 38 42 25
31 35 28

● Use analysis of variance technique and determine whether the machines

are significantly different in the mean output at level of significance 5%.

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One- way ANOVA

Consider a experiment involving three drugs, say A, B and C for

comparison. Each of these drugs are applied on 5 patients. Let the responses
to these are as given below.
Reaction time(in hrs.)

Patients
Drugs 1 2 3 4 5 Total Mean
A 7 5 6 3 4
B 10 12 9 8 6
C 14 16 13 12 10
GT GT
Mean

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Steps
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e) Find the sum of squares within the class or sum of squares due to
error (SSE):

SSE = TSS – SSTr

SSE = 210 – 160

= 50

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Source of Variation DF Sum of Mean F ratio F-Table

Squares sum (F cal)
(SS) Squares
(MSS)
Between treatments 3-1 = 2 160 80

Within treatments ( 15 -3 = 12 50 4.17 (80 / 4.17)= 6.93

Experimetal error) 19.184
Total 15-1 = 14 210

4). Critical value of F or Table value of F:

The table value is obtained from F-table for (k-1, n-k) df at α % & denoted it as F tab

F(2,12) at α=0.01
6.93

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5. Decision criteria:

If Fcal ≥ Ftab, ⇒ Reject Ho and concluded that the class means or

treatment means are significantly different ( i.e. class means are not
same).
19.184 > 6.93

There is a significant difference in mean reaction time among drugs

(i.e. mean reaction time is not same for different drugs)

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Least Significant Difference Test (LSD) or

Critical Difference (CD)
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Post Hoc Tests:

1. Least significant difference (LSD) or Critical

difference(CD)
2. Tukey’s HSD Test
3. Student-Newman-Kuel Test
4. Duncan’s multiple range Test (DMRT)
5. Bonferroni Test
6. Games-Howell test

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Tukey’s HSD Test :

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Tukey’s HSD Test :

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Statistical models for experimental designs:

●

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Fixed Effects:
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(Analysis of variance model or model-I)

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Random effect model:

(Component of variance model or model II)
●

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In random effects model, the main emphasis is on estimating and testing

the variability among the effects of different treatments.

NH: σ2= 0
AH: σ2 ǂ 0

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Two-way Classification: (Two-way ANOVA):

Let us consider the case when there are two factors which may affect the
variate yij values under study.

Ex: The yield of cow milk may be affected by rations (feeds) as well as
the varieties (breeds) of the cows.

Let us now suppose that the n cows are divided into ‘h’ different groups
or classes according to their breed, each group containing ‘k’ cows and
then let us consider the effect of k treatments (rations) given at random
to cows in each group on the yield of milk.

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Let the suffix ‘i’ refer to the treatments (rations/feeds) and

‘j’ refer to the varieties (breed of the cow), then the yields
of milk is yij (i:1,2, …..k); j:(1,2….h) of n (= RXC) cows
furnish the data for the comparison of the treatments
(rations) as well as varieties.

The yields may be expressed as variate values in the

following k X h two way table.

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Rations Breeds Total Mean

1 2 3 j h
1

Total Grand
Total
(GT)
Mean

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The total variation in the observation yij can be split into the following three
components:
(i) The variation between the treatments (rations)
(ii) The variation between the varieties (breeds)
(iii) The inherent variation within the observations of treatments and
varieties.
The first two types of variations are due to assignable causes which can be
detected and controlled by human endeavour and the third type of variation
due to chance causes which are beyond the control of human hand.

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Test procedure for two -way analysis: The steps involved in carrying out
the analysis are:

1(a) . Null hypothesis (Ho):

Ho : μ1. = μ2. = ……= μk. (for comparison of treatment/ rations) i.e., there is
no significant difference between rations (treatments)
H1 : There is a significant difference between rations (treatments)

1(b).
H0: μ.1 = μ.2 = … =μ.h (for comparison of varieties/ breed) i.e. there is no
significant difference between varieties ( breeds).
H1: There is a significant difference between varieties ( breeds)

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ANOVA TABLE

Sources of Variation d.f. Sum of M.S.S F ratio

squares (S.S.)

Between Treatments (Feeds) k-1 SSTr MST= SSTr / k-1 FT =MST / MSE

Between Varieties (Breeds) h-1 SSVt MSV=SSVt / h-1 FV =MSV / MSE

Within treatment and (k-1)(h-1) SSE MSE= SSE /

varieties (k-1)(h-1)
(Error)

Total n-1 TSS

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4) Critical values of F table (Ftab):

(i) For comparison between treatments, obtain F-table value for [k-1, (k-1)
(h-1)] df at a level of significance and denoted it as Ftab.

(ii) For comparison between Varieties, obtain F-table value for [k-1, (k-1)
(h-1)] df at a level of significance and denoted it as Ftab.

5) Decision criteria.

(i) If FCalT ≥ Ftab for [k-1, (k-1) (h-1)] df at a level of significance, H0 is

rejected.

(ii) If Fcal V ≥ Ftab for [h-1, (k-1) (h-1)] df at a level of significance, H1 is

rejected.

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Example:
●

Methods
M1 M2 M3
1 7.5 7.0 7.1
Analyst 2 7.4 7.2 6.7
3 7.3 7.0 6.9
4 7.6 7.2 6.8
5 7.4 7.1 6.9

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Methods
M1 M2 M3 Row Total, Ti.

(Analyst)
1 7.5, 7.0 7.1 21.6 466.56
7.4,6.8
Analyst 2 7.4 7.2 6.7 21.3 453.69

3 7.3 7.0 6.9 21.2 449.44

4 7.6 7.2 6.8 21.6 466.56

5 7.4 7.1 6.9 21.4 457.96

Column Total, 37.2 35.5 34.4 GT=107.1 2294.21
T.j

(Methods)

1383.84 1260.25 1183.36 3827.45

Means 7.44 7.10 6.88

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Steps:
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SS due to Error = SSE = TSS - SSA - SSM

= 0.976 – 0.043 – 0.796

=0.137

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ANOVA TABLE

Sources of DF Sum of M.S.S F ratio F table

Variation squares (S.S.)

Analyst k-1 =5-1=4 SSA = 0.043 F(0.05,4,

8)
= 3.837

Methods h-1 =3-1=2 SSM=0.796 F(0.05,2,

8)
= 4.458

Error (k-1)(h-1) SSE=0.137

(4X2)=8

Total n-1 =15-1=14 TSS =0.976

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Decision criteria:

Since the calculated FA value(0.631) is < 1, Accept Null hypothesis,

Hence, there no significant difference between the analyst.

Since the calculated FB value (23.27) is greater than the F table value
(4.458), Reject Null hypothesis, Thus, we conclude that the methods
differ significantly at 5 % level of significance.

We are interested to find out which pairs of levels of methods means

differ significantly.

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Comparison of Means:

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Pairwise comparison:

Methods Means Difference between Compare with CD (0.1914)

means
1 >CD
2 >CD
3 >CD

Hence, there is significant difference between all the three pairs of

means of methods, the greatest difference being between means of
methods 1 and 3, followed by the methods 1 and 2, and finally
between the methods 2 and 3.

CD=0.1914

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Model (Two way ANOVA):

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Two-way Analysis of variance with m-observations per cell

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Statement of Hypothesis:
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ANOVA TABLE
Sources of DF Sum of M.S.S F ratio F table
Variation squares (S.S.)

Factor A p-1 SSA = FA =

Factor B q -1 SSB= FB =

Interaction (p-1)(q-1) SSI=

AB F I=

Error pq (m-1)

Total (pqm - 1) TSS =

N-1

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Statement of Hypothesis:
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SS due Error = SSE = TSS - SSA - SSB - SSI

= 13.48 – 10.34 – 0.86 – 0.566

= 1.714

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ANOVA TABLE
Sources of DF Sum of M.S.S F ratio F table
Variation squares
(S.S.)
Order of 5-1 = 4 10.342 F0.05(4,50) = @5
gravida 2.55
@1 3.71

Age group 5 -1 = 4 0.868 F0.05(4,50) =

of mother 5@ 2.55
@1 3.71
(5-1)(5-1) 0.566 F0.05(16,50)=
Interaction = 16 @5 1.85

Error 5*5 (3-1)

= 50 1.714
Total 75-1 = 74 13.48

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Conclusion:

We conclude that F ratio (F cal) for interaction is not significant. But

the order of gravida and age group of mother have significantly
different. Both affect the birth weight of babies significantly at 5 %
level of significance.

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Comparison of Means:

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Pairwise comparison:

Order of Means
gravida
1
2
3
4
5

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Order of Difference between Compare with CD (0.134)

gravida means
1 &2 >CD
1 &3 >CD
1 &4 >CD
1 &5 >CD
2 &3 >CD
2 &4 >CD
2 &5 >CD
3 &4 >CD
3 &5 >CD
4 &5 <CD

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Comparison of Means:

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Pairwise comparison:

Age group of Means

mother
1 15-20
2 20-25
3 25-30
4 30-35
5 35 and above

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Age group Difference between Compare with CD (0.134)

of mother means
1 &2 >CD
1 &3 <CD
1 &4 >CD
1 &5 <CD
2 &3 >CD
2 &4 >CD
2 &5 >CD
3 &4 >CD
3 &5 <CD
4 &5 <CD

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Critical Difference (CD)

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One-Way ANOVA

A random sample of the students in each row was taken

The score for those students on the second exam was recorded

Front: 82, 83, 97, 93, 55, 67, 53,54,58

Middle: 83, 78, 68, 61, 77, 54, 69, 51, 63
Back: 38, 59, 55, 66, 45, 52, 52, 61,60
 Basic Principles of Experimental Designs:
The purpose of designing an experiment is to increase the precision
of the experiment. In order to increase the precision, we try to
reduce the experimental error. To reduce the experimental error, we
adopt certain principles known as basic principles of experimental
design.

The basic principles of design of experiments are:

1) Replication
2) Randomization
3) Local control
 Replication:
The repeated application of the treatments under investigation is
known as replication.
If the treatment is applied only once we have no means of
knowing about the variations in the results of a treatments. Only
when we repeat the application of the treatment several times,
we can estimate the experimental error. As the number of
replications is increased the experimental error will be reduced.
Major functions/role of the replications:

1. Replication is essential to valid estimate of experimental

error.

2. Replication is used to reduce the experimental error and

increase the precision.

3. Replication is used to measure the precision of an

experiment. If replication increases precision increases.
 Randomization:
When all the treatments have equal chance of being allocated to different
experimental units it is known as randomization.
Or
Allocation of treatments to experimental units in such a way that experimental
unit has equal chance of receiving any of the treatments is called
randomization.
 Major function/role of the randomization:

1. Randomization is used to make experimental error

independent.
2. Randomization makes test valid in the analysis of experimental
data.
3. Randomization eliminates the human biases.
4. Randomization makes free from systematic influence of
environment.
Local control:

Experimental error is based on the variations in experimental

material from experimental unit to experimental unit. This suggests
that if we group the homogenous experimental units into blocks, the
experimental error will be reduced considerably. Grouping of
homogenous experimental units into blocks is known as local control
of error.
 Major function/role of local control:
1. To reduce the experimental error.
2. Make the design more efficient.
3. It makes any test of significance more sensitive and powerful.

Remarks: In order to have valid estimate of experimental error the

principles of replication and randomization are used.

In order to reduce the experimental error, the principles of

replication and local control are used.
 Completely Randomized Design (CRD):

CRD is the basic single factor design. In this design, the treatments are
assigned completely at random so that each experimental unit has the
same chance of receiving any one treatment.
But CRD is appropriate only when the experimental material is
homogeneous.
CRD is not preferred in field experiments.
In laboratory experiments, pot culture experiment and greenhouse
studies it is easy to achieve homogeneity of experimental materials and
therefore CRD is most useful in such experiments.
 Definition:
It is defined as the design in which first the field is divided into a
number of experimental units (small plots) depending upon the number
of treatments and number of replications for each treatment, and then
treatments are assigned completely at random so that each experimental
unit has the same chance of receiving any one treatment.
(It is also known as non-restrictional design)
 Layout of CRD:
Completely randomized design is the one in which all the experimental units are
taken in a single group which are homogeneous as far as possible. The
randomization procedure for allotting the treatments to various units will be as
follows.
1. Determine the total number of experimental units.
2. Assign a plot number to each of the experimental units starting from left to right
for all rows.
3. Assign the treatments to the experimental units by using random numbers.
•
1 2 3 4
8 7 6 5
9 10 11 12
16 15 14 13
17 18 19 20
•Then ‘n’ distinct three-digit random numbers are selected
from the random number table.

•The random numbers are written in order and are ranked.

•The lowest random number is given as rank 1 and the highest

rank is allotted to the largest number.

•These ranks correspond to the plot number, the first set of ‘r’
units are allocated to treatment t1, the next ‘r’ units are
allocated to treatment t2 and so on. This procedure is
continued until all treatments have been applied.
Random Rank Treatment to be
Number applied
807 18 t 1 t1
186 4 t1 t1
410 10 t1
345 9 (r times)
626 14 (5 times)
340 7 t2 t2
883 19 t2 t2
569 13 t2
341 8 (r times)
094 2 (5 times)
322 6 t3 t3
252 5 t3 t3 (r times)
047 1 t3
469 12 (5 times)
632 15
183 3 t4 t4
417 11 t4 t4
782 17 t4
969 20 (r times)
697 16 (5 times)
Note: Only replication and randomization principles are adopted in this
design. But local control is not adopted (because experimental material
is homogeneous).
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The Analysis of Variance (ANOVA) model for CRD:

●

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Classes / Total Mean

Groups
1 ……..

2 ……..

3 ……..

. . . . ….
. . . . …..
k ……..

Grand
Total
(GT)

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Test Procedure: The steps involved in carrying out the analysis are:

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ANOVA Table

Source of Variation DF Sum of Mean F ratio

Squares Squares
Treatments k-1 SSTr

Error n-k ESS

Total n-1 TSS

4) Critical value of F or Table value of F:

The table value is obtained from F-table for (k-1, n-k) df at α % & denoted it as F tab

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Advantages of CRD:

1. layout is very easy.

2. There is complete flexibility in this design i.e. any number of treatments and replications for each
treatment can be tried.

3.Whole experimental material can be utilized in this design

4.This design yields maximum degrees of freedom for experimental error.

5.The analysis of data is simplest as compared to any other design.

6.Even if some values are missing the analysis will be remains simple.

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Disadvantages of CRD :

1. It is difficult to find homogeneous experimental units in all respects and hence CRD is seldom
suitable for field experiments as compared to other experimental designs.

2. It is less accurate than other designs.

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Uses of CRD:
CRD is more useful under the following circumstances.

1) When the experimental material is homogeneous i.e., laboratory, or green house, playhouses, pot
culture etc.

2) When the quantity or amount of experimental material of any one or more of the treatment is
limited or small.

3) When there is a possibility of any one or more observations or experimental unit being destroyed.

4) In small experiments where there is a small number of degrees of freedom.

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Randomized Complete Block Design (RCBD)

Situation to adopt RCBD:

When the experimental material is heterogeneous, the experimental material is

grouped into homogenous sub-groups called blocks. As each block consists of
the entire set of treatments and number of blocks is equivalent to number of
replications.

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Definition:

In RCBD, first heterogeneous experimental material (units) is divided into homogenous

material (units) called blocks, such that the variability within blocks is less than the
variability between blocks. The number of blocks is chosen to be equal to the number of
replications for the treatments and each block consists of as many experimental units as the
number of treatments (i.e. each block contains all treatments). Then the treatments are
allocated randomly to the experimental units within each block freshly and independently
such a way that each treatment appears only once in a block. This design is also known as
Randomized Block Design (RBD).

(This design is also known as Randomized Complete Block Design - RBD)

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Layout of RCBD

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● Let r = 4, t = 3

Low ----fertility--- High

Block
I
Block
II
Block
II
Block
IV
t2 t1 t3 t1
t1 t2 t1 t2
Field t3 t3 t2 t3

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Allocation to Block I and II

Rand. No Rank Treatment B1 B2 B3 B4

755 2 T1 T2
003 1 T2
T1
812 3 T3
T3

B1 B2 B3 B4
Rand. No Rank Treatment
T2 T1
053 1 T1
T1 T2
684 2 T2
749 3 T3 T3 T3

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Allocation to Block III and IV

Rand. No Rank Treatment B1 B2 B3 B4

T2 T1 T3
215 2 T1
T1 T2 T1
396 3 T2
199 1 T3 T3 T3 T2

B1 B2 B3 B4
Rand. No Rank Treatment T2 T1 T3 T1
178 1 T1
T1 T2 T1 T2
310 2 T2
976 3 T3 T3 T3 T2 T3

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Analysis of Variance (ANOVA) model for RCBD

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Arrangement of results for analysis

1 2 ….….…j……….. r Total
1 y11 y21 y12 y22 ………………. y1r T1
2 . . ………………. y2r . T2
.i .
. . ………………. .
. yt1 . yt2 ………………. . *"
………………. .ytr .
………………… Tt
.
T

Total R1 R2 …… ………. Rr GT

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ANOVA Table

Source of Variation df Sum of Mean

Squares Squares
Between Replications / r-1 RSS RMS
Block
Between Treatments t-1 TSS TMS
Within treatments (r-1) (t-1) ESS EMS
(Error)

Total n-1 TSS

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Test Procedure:

1. Null hypothesis:
2. Level of significance (α ):
3. Test Statistic:
4. calculated F statistic and F table values
5. Decision criteria

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A hardness testing machine operates by pressing a tip into a metal test “coupon.” The hardness of the
coupon can be determined from the depth of the resulting depression. Four tip types are being tested
to see if they produce significantly different readings. The coupons might differ slightly in their
hardness (for example, if they are taken from ingots produced in different heats).Thus coupon is can
be treated as a blocking factor.

Test Coupon
1 2 3 4
1 9.3 9.4 9.6 10
Type
2 9.4 9.3 9.8 9.9
of Tip
3 9.2 9.4 9.5 9.7

4 9.7 9.6 10 10.2

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Efficiency of Blocking

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Missing Plot Technique

Due to accident, mishandling, attack of pests (for agricultural experiments) or some other reason
one or more observations from an experiment may be lost or the value may be suspicious so that it
is wise to treat it as absent.

Yates (1937) considered a method of estimating the missing values, inserting the estimates and
analysing the data. This technique gives results identical with those obtained by the correct
procedure. The theory has been developed under the assumption that all the treatment contrasts are
estimable under missing value

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Replication Total
I II III IV
1 22.9 25.9 39.1 33.9
2 29.5 30.4 X 29.6
3 28.8 24.4 32.1 28.6
4 47 40.9 12.8 32.1
5 28.9 20.4 21.1 31.8
Total

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Replication Total
I II III IV
1 22.9 25.9 39.1 33.9 121.8
2 29.5 30.4 X 29.6 89.5
3 28.8 24.4 32.1 28.6 113.9
4 47 40.9 12.8 32.1 132.8
5 28.9 20.4 21.1 31.8 102.2
Total 157.1 142 105.1 156 560.2

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X= 25.64

SOV DF SSQ MSQ F ratio F Table

Replication 3 94.246 31.415 0.39 3.587

Treatment( 4 125.716 31.429 0.39 3.356

Cord)
Error 12-1=11 877.883 79.80
Total 18

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Grain yield per plant (grams) of maize of nine varieties in a randomized block
design were as tabulated below.
Varieties Rep I Rep II Rep III Total
V1 21 20 19.5
V2 19 18 18.5
V3 18.5 18 18.9
V4 27.5 X 27
V5 31 32.5 32.6
V6 31.5 30.5 32
V7 25.3 25.5 26.6
V8 39 40 38.5
V9 39 38.5 40
Total

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Varieties Rep I Rep II Rep III Total

V1 21 20 19.5 60.5
V2 19 18 18.5 55.5
V3 18.5 18 18.9 55.4
V4 27.5 X 27 54.5
V5 31 32.5 32.6 96.1
V6 31.5 30.5 32 94
V7 25.3 25 26.6 76.9
V8 39 40 38.5 117.5
V9 39 38.5 40 117.5
Total 251.8 222.5 253.6 727.9

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X= 26.9
B=0.77
CF=21100.85
Total SS= 1585.63
Replication SS= 0.99
Treat SS= 1576.80
Corrected Treat SS=1576.03
Error SS= 8.61

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SOV DF SSQ MSQ F ratio F Table

Replication 2 0.99 0.495 0.865 3.68

Treatment 8 1576.03 197 343.61 2.64

Error 15 8.61 0.574

Total 18 1585.63

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Treatment R1 R2 R3
1 14.5 14 14
2 16.5 16.9 16.7
3 X 16.7 17.4
4 17.6 16.9 17.5
5 18.5 17.9 17.6
6 19.3 18.3 18.8
7 19.5 19 X

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Treatment R1 R2 R3 Total
1 14.5 14 14 42.5
2 16.5 16.9 16.7 50.1
3 X31 16.7 17.4 34.1/2=17.05
4 17.6 16.9 17.5 52
5 18.5 17.9 17.6 54
6 19.3 18.3 18.8 56.4
7 19.5 19 X73 (19.21) 38.5
105.9/6=17.
Total 119.7 102 327.6+19.21= 346.8
65

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Treatment R1 R2 R3 Total
1 14.5 14 14 42.5
2 16.5 16.9 16.7 50.1
3 17.5 16.7 17.4 51.6
4 17.6 16.9 17.5 52
5 18.5 17.9 17.6 54
6 19.3 18.3 18.8 56.4
7 19.5 19 19.2 57.7
Total 123.4 119.7 121.2 364.3

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Bias(X31) = 0.019

Bias(X73) = 4.140

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CF=
Tss=
TrSS=

Corrected TRSS= TrSS – B1- B2

Ess

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● T3-T7
● T3-----0+1+0.5=1.5 T7=0.5+1+0 =1.5
Treatment R1 R2 R3 Total
1 14.5 14 14 42.5
2 16.5 16.9 16.7 50.1
3 17.5 16.7 17.4 51.6
4 17.6 16.9 17.5 52
5 18.5 17.9 17.6 54
6 19.3 18.3 18.8 56.4
7 19.5 19 19.2 57.7
Total 123.4 119.7 121.2 364.3

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Procedure for estimating the two missing values

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2. Enter all initial values assigned in step1, in the table of observed values and estimate the
one remaining missing observation by using the appropriate missing data formula.
(x73=19.21)
3. Enter the estimate of the missing data obtained in step 2, in the table consisting of all
observed values and the initial value (or values) assigned in step1.

4. Remove one initial value. Treat the removed value as the missing data, and estimate it
following the same missing data formula technique used in step 2.

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Repeat the foregoing procedure for the third missing observation, then for the fourth missing
observation, and so on, until all missing data have been estimated once through the missing data
formula

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Data Transformation in DOE

▪ Violation of assumptions of ANOVA may result in unreliable statistical tests and the
unacceptability of the conclusions.

▪ A solution is often to 'transform' the data to conform to a normal probability distribution. For this,
we take the original data apply a formula and carry out ANOVA on the transformed data.

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1. Logarithmic transformation:

● Most popular among the different types of transformations used to transform skewed data
to approximately conform to normality. If the original data follows a non–normal
distribution, so, then the log-transformed data follows a normal or near normal distribution.

● When the original observation Y is converted to log Y, the conversion is known as log
transformation.

● Log to base 10 is commonly used.

● The logarithmic transformation is mostly appropriate for data where the standard deviation is
proportional to the mean.

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2. Square root transformation:

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Angular transformation (Arc Sine) / (Inverse Sine)

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Advantages of RBD

All the three principles of design of experiments are used, the conclusions drawn from RBD are
more valid and reliable.

If data from individual units be missing then, analysis can be done by estimating it.

Most popular design in view of its simplicity, flexibility and validity.

RCBD has been shown to be more efficient or accurate than CRD. The elimination of block sum
of squares from error sum of squares, usually results in a decrease of error sum of squares.

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Disadvantages of RBD

In field experiments, it is usually observed that as the number of treatments increases, the block
size increases and so one has lesser control over error.

It cannot control two sided variation of experimental material simultaneously. That is why, it is not
recommended when experimental material contains considerable variability.

If replication is not same, the only remedy is to adopt CRD

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LATIN SQUARE DESIGN (LSD)

● When the experimental material is divided into rows and columns and the
treatments are allocated such that each treatment occurs only once in a row
and once in a column, the design is known as Latin Square Design (LSD).

● In LSD the number of rows and columns are equal. Hence the arrangement
will form a square. It is identified as 5X5 Latin squares, 6X6 Latin squares,
etc.

● In LSD each row and each column is a complete block or replication

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The major feature of this design is its capacity to simultaneously handle two
known sources of variation among experimental units.

Two directional blocking in a LS Design, commonly referred to as row-blocking

and column-blocking.

This procedure makes it possible to estimate the variation among row-blocks as

well as column blocks and to remove them from experimental error.

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A Latin Square experiment is assumed to be a three-factor experiment.

The factors are rows, columns and treatments.

It is assumed that there is no interaction between rows, columns and

treatments.

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Example

A courier company is interested in deciding between five brands

(D,P,F,C and R) of car for its next purchase of fleet cars. The company
wants to carry out a study that will enable them to compare the brands with
respect to operating costs. For this purpose they select five drivers (Rows). In
addition, the study will be carried out over a five week period (Columns =
weeks).

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● An animal scientists wishes to study weight gain in piglets but knows that
both litter membership and initial weights significantly affect the response.

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Latin Square Designs

Selected Latin Squares can be in these forms:

A B C
B C A
C D A

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4x4

A B C D A B C D A B C D A B C D

B C D A B A D C B C D A B D A C

C D A B C D B A C D A B C A D B

D A B C D C A B D A B C D C B A

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5x5
A B C D E
B A E C D
C D A E B
D E B A C
E C D B A

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A B C D E F
B F D C A E
C D E F B A
D A F E C D
E C A B F D
F E B A D C

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Randomization and Layout

The process of randomization and layout for a LS design is shown below for an experiment with
five treatments A, B, C, D, and E.

Step1: Select a sample LS plan with five treatments.

Example: 5 x 5 Latin square plan

A B C D E
B A E C D
C D A E B
D E B A C
E C D B A
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Step 2: Randomize the row arrangement of the plan selected in step 1, following one of the
randomization schemes.

Select five three-digit random numbers from random number table; for example, 628, 846, 475, 902,
and 452.
Rank the selected random numbers from lowest to highest:

Random No Sequence Rank

628 1 3
846 2 4
475 3 2
902 4 5
452 5 1

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Step 3 : Use the rank to represent the existing row number of the selected plan and the sequence to
represent the row number of the new plan. For our example, the third row of the selected plan
(rank = 3) becomes the first row (sequence =1) of the new plan; the fourth row of the selected plan
becomes the second row of the new plan; and so on. The new plan, after the row randomization is:

C D A E B
D E B A C
B A E C D
E C D B A
A B C D E

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Step 4. Randomize the column arrangement, using the same procedure used for row arrangement in
step 2. For our example, the five random numbers selected and their ranks are:

Random No Sequence Rank

792 1 4
032 2 1
947 3 5
293 4 3
196 5 2

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The rank will now be used to represent the column number of the plan obtained in step 2 (i.e., with
rearranged rows) and the sequence will be used to represent the column number of the final plan. For
our example, the fourth column of the plan obtained in step 2 becomes the first column of the final
plan, the first column of the plan of step 2 becomes the second column of the final plan, and so on. The
final plan, which becomes the layout of the experiment is:

E C B A D
A D C B E
C B D E A
B E A D C
D A E C B

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Grain Yield of four promising maize hybrids (A, B, C and D) from an experiment
with Latin Square Design

Col 1 Col 2 Col 3 Col 4 Total Mean

Row 1 1.640 (B) 1.210 (D) 1.425 (C) 1.345 (A)
Row 2 1.475 (C) 1.185 (A) 1.400 (D) 1.290 (B)
Row 3 1.670 (A) 0.710 (C) 1.665 (B) 1.180 (D)
Row 4 1.565 (D) 1.290 (B) 1.655 (A) 0.660 (C)
Total
Mean

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ANOVA

SOV DF SSQ MSQ F ratio F Table

Row (t-1)=3 0.0301 0.0100
Column (t-1)=3 0.8273 0.2757 12.82 4.76
Treatment (t-1) =3 0.4268 0.1422 6.59 4.76
Error (t-1)(t-2)=6 0.1295 0.0215

Total 15 1.413

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● CD= ?

Pairs Diff CD=0.254

Treatment Mean A-B -0.007
A 1.464 A-C 0.396
B 1.471 A-D 0.125
C 1.068 B-C 0.403
D 1.339 B-D 0.132
C-D -0.271

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Efficiency of LSD

In estimating the efficiency of LSD over RCBD, we have to consider the type of blocks. If the
LSD had been RCBD with columns as blocks it is termed as column blocking,

similarly, if the LSD had been RCBD with rows as blocks termed as row blocking.

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When the error df is less 20, the relative efficiency has to be adjusted by multiplying the
precision factor

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Advantages of LSD
Since total variation is divided into three parts namely rows, columns and
treatments, the error variance is reduced considerably. It happens due to
the fact that rows and columns being perpendicular to each other,
eliminates the two-way heterogeneity up to a maximum extent.
LSD is more efficient than RBD or CRD.

When missing values are present, missing plot technique can be used and
analysed.

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Disadvantages of LSD
This design is not as flexible as RBD or CRD as the number of treatments is
limited to the number of rows and columns.

LSD is seldom used when the number of treatments is more than 12.

LSD is not suitable for treatments less than five. Because of the limitations
on the number of treatments, LSD is not widely used in agricultural
experiments.

Note: The number of sources of variation is two for CRD, three for RBD
and four for LSD.

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Factorial Experiment
When two or more number of factors are investigated simultaneously in a single
experiment such experiments are called as factorial experiments.
The experiment, in which the effect of a number of levels of a factor is to be
assessed in combination with levels of other factor(s) simultaneously are called as
factorial experiment.
The name factorial experiment was given by F.Yates in 1926
.

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Terminologies

Factor: Factor refers to a set of related treatments. We may apply of

different doses of nitrogen to a crop. Hence nitrogen irrespective of doses is
a factor.

Levels of a factor: Different states or components making up a factor are

known as the levels of that factor. Eg: different doses of nitrogen
different doses of drug

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Types of factorial Experiment

●

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2. Asymmetrical factorial experiment:

If different factors have the different number of levels in the experiment is
known as asymmetrical factorial experiment. it is called as mixed factorial.

For example, if there are 3 varieties v0 , v1 and v2 and 2 dates of sowing d0

and d1 the treatment combinations will be vodo, vod1, v1do, v1d1, v2do v2d1.

Which is also called as 3X2 factorial experiment

For varying number of levels the arrangement is described by the product.

For example, an experiment with 3 factors each at 2 levels, 3 levels and 4
levels respectively then it is known as 2 X 3 X 4 factorial experiment.

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Simple and Main Effects

● Simple effect of a factor is the difference between its responses for a
fixed level of other factors.

● Main effect is defined as the average of the simple effects.

● Interaction is defined as the dependence of factors in their responses.

Interaction is measured as the mean of the differences between simple
effects.

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Advantages of Factorial Experiment:

● We have opportunity to evaluate the combined effect of two or more factors with
different levels simultaneously in single experiment.

● We can study individual effects of each factor as well as their interactions

● Information obtained from factorial experiment is much more than that obtained from
series of single factor experiments

● Factorial experiment results in considerable saving of experimental resources.

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Disadvantages of Factorial Experiments:

❑ When the number of factors or levels of factors or both are increased, the number of
treatment combinations will increases. Consequently, the block size will increase as result
difficult to ensure homogeneity of experimental material. This will leads to increase in
experimental error and loss of precision in the experiment.

❑ Sometimes research may not interested in certain treatment combinations. However, we

are forced to incorporate them in experiments. This leads wastage of experimental
material and resource.

❑ When several treatment combination are involved, the execution of experiment and
statistical analysis become more complex.

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Confounding in Factorial Experiments.

Confounding:
Confounding may defined as the technique of reducing the size of a replication over a
number of blocks at the cost of losing some information on the some effect which is not of
much practical important.

The process by which unimportant comparison are deliberately confused or mixed up or

entangled with the block comparisons, for the purpose of assessing more important
comparison with greater precision is called Confounding.

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Complete Confounding:
If the same effect is confounded in all the replications, then, it is know as
complete confounding.

Ex:
The confounded effect is generally of little or no value to the experimenter

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Partial Confounding:
The effect which is confounded in one replicate is being estimated and
tested on the basis of the replicates in which it is not confounded. This
system of confounding is known as partial confounding.

Ex:
AC confounded in Replication I
BC confounded in Replication II
etc

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Balanced Partial Confounding:

When all effects of the same order are confounded with incomplete blocks
equal number of times, the confounding is said to be balanced partial
confounding.

Ex:
AB, AC and BC are confounded in I, II and III replication respectively and
there are three such sets of replications.

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Unbalanced Partial Confounding:

If the effects of certain order are confounded an unequal number of times in the
replications, this system of confounding is known as unbalanced partial confounding.

EX:
AC confounded 2 times
BC confounded 3 times
AB confounded 1 time

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