Biostatistik – HPM FK UGM

Lecture 12
Multisample inference:
Analysis of Variance

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Learni ng Obj ecti ves
1.
2.
3.
4.

Describe Analysis of Variance
(ANOVA)

Explain the Rationale of ANOVA
Compare Experimental Designs
Test the Equality of 2 or More Means
a. Completely Randomized Design
b. Randomized Block Design
c. Factorial Design

5.

Use of Computer Program

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Anal ysi s of Vari ance
A analysis of variance is a technique
that partitions the total sum of squares
of deviations of the observations about
their mean into portions associated
with independent variables in the
experiment and a portion associated
with error
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Anal ysi s of Vari ance
The ANOVA table was previously

discussed in the context of regression
models with quantitative independent
variables, in this chapter the focus will
be on nominal independent variables
(factors)
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Anal ysi s of Vari ance
A factor refers to a categorical
quantity under examination in an
experiment as a possible cause of
variation in the response variable.


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Anal ysi s of Vari ance
Levels refer to the categories,
measurements, or strata of a factor
of interest in the experiment.

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Types of Experi mental Desi gns
Experimental
Designs
Completely
Randomized

Randomized
Block

One-Way
Anova

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Factorial


Two-Way
Anova

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Compl etel y Randomi zed
Desi gn

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Compl etel y Randomi zed Desi gn
1.

Experimental Units (Subjects) Are
Assigned Randomly to Treatments


2.

One Factor or Independent Variable


3.

Subjects are Assumed Homogeneous

2 or More Treatment Levels or groups


Analyzed by One-Way ANOVA

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One-Way ANOVA F-Test
1.

Tests the Equality of 2 or More (p)
Population Means

2.


Variables



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One Nominal Independent Variable
One Continuous Dependent Variable

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One-Way ANOVA F-Test
Assumpti ons
1.


Randomness & Independence of
Errors

2.

Normality


3.

Populations (for each condition) are
Normally Distributed

Homogeneity of Variance


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Populations (for each condition) have
Equal Variances
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One-Way ANOVA F-Test Hypotheses
H0: 1 = 2 = 3 = ... = p




All Population Means
are Equal
No Treatment Effect

Ha: Not All j Are Equal





At Least 1 Pop. Mean
is Different
Treatment Effect
 1  2  ...  p

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One-Way ANOVA F-Test Hypotheses
H0: 1 = 2 = 3 = ... = p




All Population Means
are Equal
No Treatment Effect

f(X)

1 = 2 = 3

Ha: Not All j Are Equal






At Least 1 Pop. Mean is
Different
Treatment Effect
 1 = 2 = ... = p
Or i ≠ j for some i, j.

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X

f(X)

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1 =  2  3

X
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One-Way ANOVA
Basi c Idea
1.

Compares 2 Types of Variation to Test
Equality of Means

2.

If Treatment Variation Is Significantly
Greater Than Random Variation then
Means Are Not Equal

3.

Variation Measures Are Obtained by
‘Partitioning’ Total Variation

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One-Way ANOVA
Parti ti ons Total Vari ati on

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One-Way ANOVA
Parti ti ons Total Vari ati on
Total variation

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One-Way ANOVA
Parti ti ons Total Vari ati on
Total variation

Variation due to
treatment

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One-Way ANOVA
Parti ti ons Total Vari ati on
Total variation

Variation due to
treatment

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Variation due to
random sampling

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One-Way ANOVA
Parti ti ons Total Vari ati on
Total variation

Variation due to
treatment

Variation due to
random sampling

Sum of Squares Among
Sum of Squares Between
Sum of Squares
Treatment
Among Groups Variation
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One-Way ANOVA
Parti ti ons Total Vari ati on
Total variation

Variation due to
treatment

Variation due to
random sampling

Sum of Squares Among
Sum of Squares Between
Sum of Squares
Treatment (SST)
Among Groups Variation
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Sum of Squares Within
Sum of Squares Error
(SSE)
Within Groups Variation

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Total Vari ati on
SS Total   Y11  Y   Y21  Y     Yij  Y 
2

2

2

Response, Y

Y

Group 1
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Group 2
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Group 3
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Treatment Vari ati on
SST  n1 Y1  Y   n2 Y2  Y     n p Y p  Y 
2

2

2

Response, Y

Y3
Y
Y1
Group 1
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Y2
Group 2

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Group 3
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Random (Error) Vari ati on

SSE  Y11  Y1   Y21  Y1     Y pj  Y p 
2

2

2

Response, Y

Y3
Y1
Group 1
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Y2

Group 2
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Group 3
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One-Way ANOVA F-Test
Test Stati sti c
 1.


Test Statistic
F = MST / MSE

STT /  p  1

SSE / n  p 

• MST Is Mean Square for Treatment
• MSE Is Mean Square for Error

 2.



Degrees of Freedom
1 = p -1
2 = n - p
• p = # Populations, Groups, or Levels
• n = Total Sample Size

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One-Way ANOVA
Summary Tabl e
Source of Degrees Sum of
Variation
of
Squares
Freedom

Mean
Square
(Variance)

Treatment

p-1

SST

MST =
MST
SST/(p - 1) MSE

Error

n-p

SSE

MSE =
SSE/(n - p)

Total

n-1

SS(Total) =
SST+SSE

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F

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One-Way ANOVA F-Test Cri ti cal
Val ue
If means are equal,
F = MST / MSE  1.
Only reject large F!

Reject H0
Do Not
Reject H0
0



Fa ( p1, n p)

F

Always One-Tail!
© 1984-1994 T/Maker Co.

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One-Way ANOVA F-Test Exampl e
Food1 Food2 Food3
As a vet epidemiologist
25.40 23.40 20.00
you want to see if 3 food
26.31 21.80 22.20
supplements have
different mean milk yields. 24.10 23.50 19.75
You assign 15 cows, 5 per 23.74 22.75 20.60
25.10 21.60 20.40
food supplement.
Question: At the .05 level,
is there a difference in
mean yields?
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One-Way ANOVA F-Test
Sol uti on






H0:  1 =  2 =  3
Ha: Not All Equal
 = .05
1 = 2 2 = 12
Critical Value(s):
 = .05

0
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3.89

F

Test Statistic:

F

MST
MSE



23.5820
.9211

 25.6

Decision:
Reject at  = .05
Conclusion:
There Is Evidence Pop.
Means Are Different

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Summary Tabl e
Sol uti on
Source of Degrees of Sum of
Variation Freedom Squares
Food

3-1=2

47.1640

Error

15 - 3 = 12 11.0532

Total

15 - 1 = 14 58.2172

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Mean
Square
(Variance)

F

23.5820

25.60

.9211

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SAS CODES FOR ANOVA











Data Anova;
input group$ milk @@;
cards;
food1 25.40
food2 23.40
food1 26.31
food2 21.80
food1 24.10
food2 23.50
food1 23.74
food2 22.75
food1 25.10
food2 21.60
;
run;

food3 20.00
food3 22.20
food3 19.75
food3 20.60
food3 20.40

proc anova; /* or PROC GLM */
 class group;
 model milk=group;
 run;


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OUTPUT - ANOVA
Source

DF

Model

2

Error

12

Corrected Total

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Sum of
Squares

Mean Square

F Value

Pr > F

47.16400000

23.58200000

25.60

F
0.0030

resp Mean
43350.00

Source

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F Value

F Value

Pr > F

11.99 0.0006
1.90 0.1759

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SAS OUTPUT - LSD
Means with the same letter are not significantly
different.
t Grouping

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Mean

N

trt

A

50200

5

C:

B
B
C B
C
C

44800

5

D:

41000

5

A:

37400

5

B:

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SAS OUTPUT - Bonferroni
Means with the same letter are not significantly
different.
Bon Grouping
A
A
B A
B
B C
C
C
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Mean

N

trt

50200

5

C:

44800

5

D:

41000

5

A:

37400

5

B:

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Factori al Experi ments

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Factori al Desi gn
 1.

Experimental Units (Subjects) Are
Assigned Randomly to Treatments


Subjects are Assumed Homogeneous

 2.

Two or More Factors or
Independent Variables


Each Has 2 or More Treatments (Levels)

 3.

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Analyzed by Two-Way ANOVA
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Advantages
of Factori al Desi gns
1.

Saves Time & Effort
e.g., Could Use Separate Completely
Randomized Designs for Each Variable

2.

Controls Confounding Effects by
Putting Other Variables into Model

3.

Can Explore Interaction Between
Variables

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Two-Way ANOVA
1.

Tests the Equality of 2 or More
Population Means When Several
Independent Variables Are Used

2.

Same Results as Separate One-Way
ANOVA on Each Variable
-

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But Interaction Can Be Tested
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Two-Way ANOVA Assumpti ons
1.

Normality
• Populations are Normally Distributed

2.

Homogeneity of Variance
• Populations have Equal Variances

3.

Independence of Errors
• Independent Random Samples are Drawn

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Two-Way ANOVA
Data Tabl e
Factor
A
1
Y111
1
Y112
Y211
2
Y212
:
:
Ya11
a
Ya12
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Factor B
2
...
Y121 ...
Y122 ...
Y221 ...
Y222 ...
:
:
Ya21 ...
Ya22 ...

b
Y1b1
Y1b2
Y2b1
YX
2b2
:
Yab1
Yab2

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Observation k

Yijk
Level i
Factor
A

Level j
Factor
B

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Universitas Gadjah Mada, Faculty of Medicine, Dept. Biostat. Epi. and Pop. Health

Two-Way ANOVA
Nul l Hypotheses
1. No Difference in Means Due to
Factor A


H0: 1. = 2. =... = a.

2. No Difference in Means Due to
Factor B


3.

H0: .1 = .2 =... = .b

No Interaction of Factors A & B


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H0: ABij = 0
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Two-Way ANOVA
Total Vari ati on Parti ti oni ng
Total Variation
SS(Total)
Variation Due to
Treatment A

Variation Due to
Treatment B

SSA

SSB



Variation Due to
Interaction

Variation Due to
Random Sampling

SS(AB)
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SSE
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Two-Way ANOVA
Summary Tabl e
Source of Degrees of Sum of
Variation
Freedom Squares

Mean
Square

F

A
(Row)

a-1

SS(A)

MS(A)

MS(A)
MSE

B
(Column)

b-1

SS(B)

MS(B)

MS(B)
MSE

AB
(a-1)(b-1)
(Interaction)
Error
Total
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n - ab

SS(AB)
SSE

MS(AB) MS(AB)
MSE

MSE
Same as
n-1
SS(Total)
Other
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Designs
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Interacti on
1. Occurs When Effects of One Factor
Vary According to Levels of Other
Factor
2. When Significant, Interpretation of
Main Effects (A & B) Is Complicated
3.

Can Be Detected




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In Data Table, Pattern of Cell Means in One
Row Differs From Another Row
In Graph of Cell Means, Lines Cross
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Graphs of Interacti on
Effects of Gender (male or female) & dietary
group (sv, lv, nor) on systolic blood pressure
Interaction

Average
Response

No Interaction

male

Average
Response

male

female
sv
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lv

nor
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female
sv

lv

nor
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Two-Way ANOVA F-Test Exampl e
Effect of diet (sv-strict vegetarians, lvlactovegetarians, nor-normal) and gender
(female, male) on systolic blood pressure.
Question: Test for interaction and main effects
at the .05 level.

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SAS CODES FOR ANOVA
data factorial;
input dietary$ sex$
sbp;
cards;
sv male 109.9
sv male 101.9
sv male 100.9
sv male 119.9
sv male 104.9
sv male 189.9
sv female 102.6
sv female 99
sv female 83 .6
sv female 99.6
sv female 102.6
sv female 112.6
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lv male 116.5
lv male 118.5
lv male 119.5
lv male 110.5
lv male 115.5
lv male 105.2
nor male 128.3
nor male 129.3
nor male 126.3
nor male 127.3
nor male 126.3
nor male 125.3
nor female 119.1
nor female 119.2
nor female 115.6
nor female 119.9
nor female 119.8
nor female 119.7
;
run;
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SAS CODES FOR ANOVA
proc glm;
class dietary sex;
model sbp=dietary sex dietary*sex;
run;
proc glm;
class dietary sex;
model sbp=dietary sex;
run;
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SAS OUTPUT - ANOVA
Dependent Variable: sbp
Sum of
Source
DF
Squares Mean Square F Value Pr > F
Model
5 2627.399667
525.479933
1.96 0.1215
Error
24 6435.215000
268.133958
Corrected Total
29 9062.614667
R-Square
0.289916

Coeff Var
14.08140

Root MSE
16.37480

sbp Mean
116.2867

Source
dietary
sex
dietary*sex

DF
Type I SS Mean Square F Value Pr > F
2
958.870500
479.435250
1.79 0.1889
1 1400.686992 1400.686992
5.22 0.0314
2
267.842175
133.921087
0.50 0.6130

Source
dietary
sex
dietary*sex

DF Type III SS Mean Square F Value Pr > F
2 1039.020874
519.510437
1.94 0.1659
1 877.982292
877.982292
3.27 0.0829
2
267.842175
133.921087
0.50 0.6130

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Li near Contrast


Linear Contrast is a linear combination of the
means of populations



Purpose: to test relationship among different
group means

L  cj j

with

c

j

0

Example: 4 populations on treatments T1, T2, T3 and T4.
Contrast T1
T2
T3
T4
relation to test
L1
L2
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1
1

0
-1/2

-1
-1/2

0
0

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μ1 - μ 3 = 0
μ1 – μ2/2 – μ3/2 = 0
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Universitas Gadjah Mada, Faculty of Medicine, Dept. Biostat. Epi. and Pop. Health

T-test for Li near Contrast (LSD)


Construct a t statistic involving k group
means. Degrees of freedom of t - test: df = n-

k.
k

To test H0:

L   c j  j  0 Construct

t

j 1

L
k

c 2j

j 1

nj

s2 

Compare with critical value t1-α/2,, n-k.
Reject H0 if |t| ≥ t1-α/2,, n-k.
SAS uses contrast statement and performs an F – test df (1, n-k);
Or estimate statement and perform a t-test df (n-k).
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T-test for Li near Contrast (Scheffe)


Construct multiple contrasts involving k
group means. Trying to search for
significant contrast
k

To test H0:

L   c j  j  0 Construct
j 1

Compare with critical value.

t

L
k

c 2j

j 1

nj

s2 

a  (k  1) Fk 1,n  k ,1
Reject H0 if |t| ≥ a

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SAS Code for contrast testi ng













proc glm;
class trt block;
model resp=trt block;
Means trt /lsd bon scheffe;
contrast 'A - B = 0' trt 1 -1 0 0 ;
contrast 'A - B/2 - C/2 = 0' trt 1 -.5 -.5 0 ;
contrast 'A - B/3 - C/3 -D/3 = 0' trt 3 -1 -1 -1 ;
contrast 'A + B - C - D = 0' trt 1 1 -1 -1 ;
lsmeans trt/stderr pdiff;
lsmeans trt/stderr pdiff adjust=scheffe; /* Scheffe's test */
lsmeans trt/stderr pdiff adjust=bon;
/* Boneferoni's test
*/
estimate ‘A - B' trt 1 -1 0 0 0;



run;

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Regressi on representati on of Anova

10/27/2017

awilopo@yahoo.com

Biostatistics I: 2017-18

73

Universitas Gadjah Mada, Faculty of Medicine, Dept. Biostat. Epi. and Pop. Health

Regressi on representati on of Anova

yij  i  eij   i  eij


One-way anova:

p

  0
i1



i

Two-way anova:
yijk  ij  eijk    i   j   ij  eijk
a


i 1



b

i

 0,   j  0,
j 1

b


j 1

ij

 0 for all i and

a


i 1

ij

 0 for all j

SAS uses a different constraint

10/27/2017

awilopo@yahoo.com

Biostatistics I: 2017-18

74

Universitas Gadjah Mada, Faculty of Medicine, Dept. Biostat. Epi. and Pop. Health

Regressi on representati on of Anova


One-way anova: Dummy variables of
factor with p levels

y  0  1 x1  2 x2  ...   p1 x p1  e
1
where xi  
0


if level i
if otherwise

This is the parameterization used by SAS

10/27/2017

awilopo@yahoo.com

Biostatistics I: 2017-18

75

Universitas Gadjah Mada, Faculty of Medicine, Dept. Biostat. Epi. and Pop. Health

Concl usi on: shoul d be abl e to
1. Recognize the applications that uses
ANOVA
2. Understand the logic of analysis of
variance.
3. Be aware of several different analysis of
variance designs and understand when to use
each one.
4. Perform a single factor hypothesis test
using analysis of variance manually and with
the aid of SAS or any statistical software.
10/27/2017
sawilopo@yahoo.com

Biostatistics I: 2017-18

76

Universitas Gadjah Mada, Faculty of Medicine, Department of Public Health

Concl usi on: shoul d be abl e to
5. Conduct and interpret post-analysis of
variance pairwise comparisons
procedures.
6. Recognize when randomized block
analysis of variance is useful and be able to
perform the randomized block analysis.
7. Perform two factor analysis of variance
tests with replications using SAS and
interpret the output.

10/27/2017
sawilopo@yahoo.com

Biostatistics I: 2017-18

77

Universitas Gadjah Mada, Faculty of Medicine, Department of Public Health

Key Terms
Between-Sample
Variation
 Completely
Randomized
Design
 Experiment-Wide
Error Rate
 Factor


10/27/2017

awilopo@yahoo.com

Levels
 One-Way Analysis
of Variance
 Total Variation
 Treatment
 Within-Sample
Variation


Biostatistics I: 2017-18

78

Universitas Gadjah Mada, Faculty of Medicine, Dept. Biostat. Epi. and Pop. Health