COURSE

STAB3143 MOLECULAR APPROACHES IN CONSERVATION

SEMESTER 1 2022/2023
SET 1

TITLE
LABORATORY REPORT ON PRACTICAL 1 – ISOZYME ANALYSIS

LECTURER
PROF. DR. KALAIVANI A/P K. NADARAJAH

NAME
ADILAH AIMAN BINTI ZAFIR KHAN (A184381)
1. For each enzyme system, determine the genotype at each locus. The information on the quartenary
structure of the active enzymes is given below to assist you to determine the genotype at each locus.
ADH-dimer; PER-monomer; SDH-monomer; SOD-monomer; -6PGD-dimer; PGM-
monomer; AAT-dimer; DIA-monomer; LAP-monomer; PGI-dimer

i) ADH-dimer
Locus Phenotype Genotype Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
A 1,1 2 2 5 3 1
B 2,2 3 3 3 1 1
C 3,3 2 0 3 1 3
Adh
D 1,2 8 10 5 5 15
E 1,3 6 5 5 10 0
F 2,3 4 5 4 5 5
Total 25 25 25 25 25

ii) PER-monomer
Locus Phenotype Genotype Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
A 1,1 3 5 2 2 1
B 2,2 1 3 3 3 1
C 3,3 1 3 2 0 3
Per
D 1,2 5 5 8 10 15
E 1,3 10 5 6 5 0
F 2,3 5 4 4 5 5
Total 25 25 25 25 25

iii) SDH-monomer
Locus Phenotype Genotype Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
A 1,1 3 2 1 5 2
B 2,2 1 3 1 3 3
C 3,3 1 0 3 3 2
Sdh
D 1,2 5 10 15 5 8
E 1,3 10 5 0 5 6
F 2,3 5 5 5 4 4
Total 25 25 25 25 25

iv) SOD-monomer
Locus Phenotype Genotype Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
A 1,1 5 15 5 5 1
Sod B 2,2 5 5 20 15 10
C 1,2 15 5 0 5 14
Total 25 25 25 25 25

v) -6PGD-dimer
Locus Phenotype Genotype Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
A 1,1 5 5 15 1 5
6Pgd B 2,2 20 5 5 10 15
C 1,2 0 15 5 14 5
Total 25 25 25 25 25
vi) PGM-monomer
Locus Phenotype Genotype Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
A 1,1 5 5 1 5 15
Pgm B 2,2 20 15 10 5 5
C 1,2 0 5 14 15 5
Total 25 25 25 25 25

vii) AAT-dimer
Genotype
Locus Phenotype Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
Loci 1 Loci 2
A 1,1 1,1 2 0 8 5 5
Aat B 1,1 2,2 3 0 2 5 6
C 1,1 1,2 20 25 15 15 14
Total 25 25 25 25 25

viii) DIA-monomer
Genotype
Locus Phenotype Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
Loci 1 Loci 2
A 1,1 1,1 2 20 5 0 5
B 1,1 2,2 3 0 5 0 5
C 1,1 1,2 5 0 0 0 0
D 2,2 1,1 2 5 1 0 2
Dia E 2,2 2,2 1 0 1 20 3
F 2,2 1,2 2 0 5 0 5
G 1,2 1,1 5 0 3 0 2
H 1,2 2,2 5 0 2 5 1
I 1,2 1,2 0 0 3 0 2
Total 25 25 25 25 25

ix) LAP-monomer
Locus Phenotype Genotype Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
A 1,1 2 20 5 0 3
B 2,2 3 0 5 0 2
C 3,3 3 0 0 0 0
D 4,4 2 2 1 0 2
E 1,2 1 0 1 10 3
Lap
F 1,3 2 0 5 0 5
G 1,4 5 0 3 0 2
I 2,3 0 0 1 0 2
J 2,4 2 1 1 5 3
K 3,4 3 2 2 5 2
Total 23 25 24 20 24

x) PGI-dimer
Genotype
Locus Phenotype Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
Loci 1 Loci 2
A 1,1 1,1 6 1 25 0 2
B 2,2 1,1 2 1 0 0 3
Pgi
C 3,3 1,1 3 2 0 2 4
D 4,4 1,1 2 3 0 3 1
 E 5,5 1,1 2 3 0 0 5
F 1,2 1,1 0 5 0 5 0
G 1,3 1,1 0 2 0 6 0
I 1,4 1,1 5 0 0 0 0
J 1,5 1,1 0 0 0 0 1
K 2,3 1,1 0 0 0 0 4
L 2,4 1,1 0 2 0 2 0
M 2,5 1,1 2 0 0 3 0
N 3,4 1,1 3 0 0 0 2
O 3,5 1,1 0 3 0 0 0
P 4,5 1,1 0 0 0 0 3
Total 25 22 25 21 25

2. Calculate the frequency of alleles at all loci for all populations.

i) ADH-dimer
Locus Allele Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
2(AA)+AB+AC 2(AA)+AB+AC 2(AA)+AB+AC 2(AA)+AB+AC 2(AA)+AB+AC
F(A) = F(A) = F(A) = F(A) = F(A) =
2N 2N 2N 2N 2N
2(2)+8+6 2(2)+10+5 2(5)+5+5 2(3)+5+10 2(1)+15+0
= = = = =
1 2(25) 2(25) 2(25) 2(25) 2(25)
18 19 20 21 17
= = = = =
50 50 50 50 50
= 0.36 = 0.38 = 0.40 = 0.42 = 0.34
2(BB)+AB+BC 2(BB)+AB+BC 2(BB)+AB+BC 2(BB)+AB+BC 2(BB)+AB+BC
F(B) = F(B) = F(B) = F(B) = F(B) =
2N 2N 2N 2N 2N
2(3)+8+4 2(3)+10+5 2(3)+5+4 2(1)+5+5 2(1)+15+5
= = = = =
Adh 2 2(25) 2(25) 2(25) 2(25) 2(25)
18 21 15 12 22
= = = = =
50 50 50 50 50
= 0.36 = 0.42 = 0.30 = 0.24 = 0.44
2(CC)+AC+BC 2(CC)+AC+BC 2(CC)+AC+BC 2(CC)+AC+BC 2(CC)+AC+BC
F(C) = F(C) = F(C) = F(C) = F(C) =
2N 2N 2N 2N 2N
2(2)+6+4 2(0)+5+5 2(3)+5+4 2(1)+10+5 2(3)+0+5
= = = = =
3 2(25) 2(25) 2(25) 2(25) 2(25)
14 10 15 17 11
= = = = =
50 50 50 50 50
= 0.28 = 0.20 = 0.30 = 0.34 = 0.22
Total 1.00 1.00 1.00 1.00 1.00

ii) PER-monomer
Locus Allele Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
2(AA)+AB+AC 2(AA)+AB+AC 2(AA)+AB+AC 2(AA)+AB+AC 2(AA)+AB+AC
F(A) = F(A) = F(A) = F(A) = F(A) =
2N 2N 2N 2N 2N
2(3)+5+10 2(5)+5+5 2(2)+8+6 2(2)+10+5 2(1)+15+0
= = = = =
1 2(25) 2(25) 2(25) 2(25) 2(25)
21 20 18 19 17
= = = = =
50 50 50 50 50
= 0.42 = 0.40 = 0.36 = 0.38 = 0.34
2(BB)+AB+BC 2(BB)+AB+BC 2(BB)+AB+BC 2(BB)+AB+BC 2(BB)+AB+BC
F(B) = F(B) = F(B) = F(B) = F(B) =
2N 2N 2N 2N 2N
2(1)+5+5 2(3)+5+4 2(3)+8+4 2(3)+10+5 2(1)+15+5
= = = = =
Per 2 2(25) 2(25) 2(25) 2(25) 2(25)
12 15 18 21 22
= = = = =
50 50 50 50 50
= 0.24 = 0.30 = 0.36 = 0.42 = 0.44
2(CC)+AC+BC 2(CC)+AC+BC 2(CC)+AC+BC 2(CC)+AC+BC 2(CC)+AC+BC
F(C) = F(C) = F(C) = F(C) = F(C) =
2N 2N 2N 2N 2N
2(1)+10+5 2(3)+5+4 2(2)+6+4 2(0)+5+5 2(3)+0+5
= = = = =
3 2(25) 2(25) 2(25) 2(25) 2(25)
17 15 14 10 11
= = = = =
50 50 50 50 50
= 0.34 = 0.30 = 0.28 = 0.20 = 0.22
Total 1.00 1.00 1.00 1.00 1.00
iii) SDH-monomer
Locus Allele Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
2(AA)+AB+AC 2(AA)+AB+AC 2(AA)+AB+AC 2(AA)+AB+AC 2(AA)+AB+AC
F(A) = F(A) = F(A) = F(A) = F(A) =
2N 2N 2N 2N 2N
2(3)+5+10 2(2)+10+5 2(1)+15+0 2(5)+5+5 2(2)+8+6
= = = = =
1 2(25) 2(25) 2(25) 2(25) 2(25)
21 19 17 20 18
= = = = =
50 50 50 50 50
= 0.42 = 0.38 = 0.34 = 0.40 = 0.36
2(BB)+AB+BC 2(BB)+AB+BC 2(BB)+AB+BC 2(BB)+AB+BC 2(BB)+AB+BC
F(B) = F(B) = F(B) = F(B) = F(B) =
2N 2N 2N 2N 2N
2(1)+5+5 2(3)+10+5 2(1)+15+5 2(3)+5+4 2(3)+8+4
= = = = =
Sdh 2 2(25) 2(25) 2(25) 2(25) 2(25)
12 21 22 15 18
= = = = =
50 50 50 50 50
= 0.24 = 0.42 = 0.44 = 0.30 = 0.36
2(CC)+AC+BC 2(CC)+AC+BC 2(CC)+AC+BC 2(CC)+AC+BC 2(CC)+AC+BC
F(C) = F(C) = F(C) = F(C) = F(C) =
2N 2N 2N 2N 2N
2(1)+10+5 2(0)+5+5 2(3)+0+5 2(3)+5+4 2(2)+6+4
= = = = =
3 2(25) 2(25) 2(25) 2(25) 2(25)
17 10 11 15 14
= = = = =
50 50 50 50 50
= 0.34 = 0.20 = 0.22 = 0.30 = 0.28
Total 1.00 1.00 1.00 1.00 1.00

iv) SOD-monomer
Locus Allele Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB
F(A) = F(A) = F(A) = F(A) = F(A) =
2N 2N 2N 2N 2N
2(5)+15 2(15)+5 2(5)+0 2(5)+5 2(1)+14
= = = = =
1 2(25) 2(25) 2(25) 2(25) 2(25)
25 35 10 15 16
= = = = =
50 50 50 50 50
= 0.50 = 0.70 = 0.20 = 0.30 = 0.32
Sod 2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB
F(B) = F(B) = F(B) = F(B) = F(B) =
2N 2N 2N 2N 2N
2(5)+15 2(5)+5 2(20)+0 2(15)+5 2(10)+14
= = = = =
2 2(25) 2(25) 2(25) 2(25) 2(25)
25 15 40 35 34
= = = = =
50 50 50 50 50
= 0.50 = 0.30 = 0.80 = 0.70 = 0.68
Total 1.00 1.00 1.00 1.00 1.00

v) -6PGD-dimer
Locus Allele Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB
F(A) = F(A) = F(A) = F(A) = F(A) =
2N 2N 2N 2N 2N
2(5)+0 2(5)+15 2(15)+5 2(1)+14 2(5)+5
= = = = =
1 2(25) 2(25) 2(25) 2(25) 2(25)
10 25 35 16 15
= = = = =
50 50 50 50 50
= 0.20 = 0.50 = 0.70 = 0.32 = 0.30
6Pgd 2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB
F(B) = F(B) = F(B) = F(B) = F(B) =
2N 2N 2N 2N 2N
2(20)+0 2(5)+15 2(5)+5 2(10)+14 2(15)+5
= = = = =
2 2(25) 2(25) 2(25) 2(25) 2(25)
40 25 15 34 35
= = = = =
50 50 50 50 50
= 0.80 = 0.50 = 0.30 = 0.68 = 0.70
Total 1.00 1.00 1.00 1.00 1.00

vi) PGM-monomer
Locus Allele Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB
F(A) = F(A) = F(A) = F(A) = F(A) =
2N 2N 2N 2N 2N
Pgm 1 2(5)+0 2(5)+5 2(1)+14 2(5)+15 2(15)+5
= = = = =
2(25) 2(25) 2(25) 2(25) 2(25)
 10 15 16 25 35
= = = = =
50 50 50 50 50
= 0.20 = 0.30 = 0.32 = 0.50 = 0.70
2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB
F(B) = F(B) = F(B) = F(B) = F(B) =
2N 2N 2N 2N 2N
2(20)+0 2(15)+5 2(10)+14 2(5)+15 2(5)+5
= = = = =
2 2(25) 2(25) 2(25) 2(25) 2(25)
40 35 34 25 15
= = = = =
50 50 50 50 50
= 0.80 = 0.70 = 0.68 = 0.50 = 0.30
Total 1.00 1.00 1.00 1.00 1.00

vii) AAT-dimer
Locus Allele Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
2(AA) 2(AA) 2(AA) 2(AA) 2(AA)
F(A) = F(A) = F(A) = F(A) = F(A) =
2N 2N 2N 2N 2N
2(2+3+20) 2(0+0+25) 2(8+2+15) 2(5+5+15) 2(5+6+14)
= = = = =
Aat – L1 1 2(25) 2(25) 2(25) 2(25) 2(25)
50 50 50 50 50
= = = = =
50 50 50 50 50
= 1.00 = 1.00 = 1.00 = 1.00 = 1.00
Total 1.00 1.00 1.00 1.00 1.00
2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB
F(A) = F(A) = F(A) = F(A) = F(A) =
2N 2N 2N 2N 2N
2(2)+20 2(0)+25 2(8)+15 2(5)+15 2(5)+14
= = = = =
1 2(25) 2(25) 2(25) 2(25) 2(25)
24 25 31 25 24
= = = = =
50 50 50 50 50
= 0.48 = 0.50 = 0.62 = 0.50 = 0.48
Aat – L2 2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB
F(B) = F(B) = F(B) = F(B) = F(B) =
2N 2N 2N 2N 2N
2(3)+20 2(0)+25 2(2)+15 2(5)+15 2(6)+14
= = = = =
2 2(25) 2(25) 2(25) 2(25) 2(25)
26 25 19 25 26
= = = = =
50 50 50 50 50
= 0.52 = 0.50 = 0.38 = 0.50 = 0.52
Total 1.00 1.00 1.00 1.00 1.00

viii) DIA-monomer
Locus Allele Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
F(A) F(A) F(A) F(A) F(A)
2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB
= = = = =
2N 2N 2N 2N 2N
2(2+3+5)+5+5+0 2(20+0+0)+0+0+0 2(5+5+0)+3+2+3 2(0+0+0)+0+5+0 2(5+5+0)+2+1+2
1 =
2(25)
=
2(25)
=
2(25)
=
2(25)
=
2(25)
30 40 28 5 25
= = = = =
50 50 50 50 50
Dia – = 0.60 = 0.80 = 0.56 = 0.10 = 0.50
L1 F(B) F(B) F(B) F(B) F(B)
2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB
= = = = =
2N 2N 2N 2N 2N
2(2+1+2)+5+5+0 2(5+0+0)+0+0+0 2(1+1+5)+3+2+3 2(0+20+0)+0+5+0 2(2+3+5)+2+1+2
2 =
2(25)
=
2(25)
=
2(25)
=
2(25)
=
2(25)
20 10 22 45 25
= = = = =
50 50 50 50 50
= 0.40 = 0.20 = 0.44 = 0.90 = 0.50
Total 1.00 1.00 1.00 1.00 1.00
F(A) F(A) F(A) F(A) F(A)
2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB 2(AA)+AB
= = = = =
2N 2N 2N 2N 2N
2(2+2+5)+5+2+0 2(20+5+0)+0+0+0 2(5+1+3)+0+5+3 2(0+0+0)+0+0+0 2(5+2+2)+0+5+2
1 =
2(25)
=
2(25)
=
2(25)
=
2(25)
=
2(25)

Dia – =
25
50
=
50
50
=
26
50
=
0
50
=
25
50
L2 = 0.50 = 1.00 = 0.52 = 0.00 = 0.50
F(B) F(B) F(B) F(B) F(B)
2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB 2(BB)+AB
2 = = = = =
2N 2N 2N 2N 2N
2(3+1+5)+5+2+0 2(0+0+0)+0+0+0 2(5+1+2)+0+5+3 2(0+20+5)+0+0+0 2(5+3+1)+0+5+2
= = = = =
2(25) 2(25) 2(25) 2(25) 2(25)
 25 0 24 50 25
= = = = =
50 50 50 50 50
= 0.50 = 0.00 = 0.48 = 1.00 = 0.50
Total 1.00 1.00 1.00 1.00 1.00

ix) LAP-monomer
Locus Allele Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
F(A) F(A) F(A) F(A) F(A)
2(AA)+AB+AC+AD 2(AA)+AB+AC+AD 2(AA)+AB+AC+AD 2(AA)+AB+AC+AD 2(AA)+AB+AC+AD
= = = = =
2N 2N 2N 2N 2N
2(2)+1+2+5 2(20)+0+0+0 2(5)+1+5+3 2(0)+10+0+0 2(3)+3+5+2
1 =
2(23)
=
2(25)
=
2(24)
=
2(20)
=
2(24)
12 40 19 10 16
= = = = =
46 50 48 40 48
= 0.26 = 0.80 = 0.40 = 0.25 = 0.33
F(B) F(B) F(B) F(B) F(B)
2(BB)+AB+BC+BD 2(BB)+AB+BC+BD 2(BB)+AB+BC+BD 2(BB)+AB+BC+BD 2(BB)+AB+BC+BD
= = = = =
2N 2N 2N 2N 2N
2(3)+1+0+2 2(0)+0+0+1 2(5)+1+1+1 2(0)+10+0+5 2(2)+3+2+3
2 =
2(23)
=
2(25)
=
2(24)
=
2(20)
=
2(24)
9 1 13 15 12
= = = = =
46 50 48 40 48
= 0.20 = 0.02 = 0.27 = 0.38 = 0.25
Lap F(C) F(C) F(C) F(C) F(C)
2(CC)+AC+BC+CD 2(CC)+AC+BC+CD 2(CC)+AC+BC+CD 2(CC)+AC+BC+CD 2(CC)+AC+BC+CD
= = = = =
2N 2N 2N 2N 2N
2(3)+2+0+3 2(0)+0+0+2 2(0)+5+1+2 2(0)+0+0+5 2(0)+5+2+2
3 =
2(23)
=
2(25)
=
2(24)
=
2(20)
=
2(24)
11 2 8 5 9
= = = = =
46 50 48 40 48
= 0.24 = 0.04 = 0.17 = 0.13 = 0.19
F(D) F(D) F(D) F(D) F(D)
2(DD)+AD+BD+CD 2(DD)+AD+BD+CD 2(DD)+AD+BD+CD 2(DD)+AD+BD+CD 2(DD)+AD+BD+CD
= = = = =
2N 2N 2N 2N 2N
2(2)+5+2+3 2(2)+0+1+2 2(1)+3+1+2 2(0)+0+5+5 2(2)+2+3+2
4 =
2(23)
=
2(25)
=
2(24)
=
2(20)
=
2(24)
14 7 8 10 11
= = = = =
46 50 48 40 48
= 0.30 = 0.14 = 0.17 = 0.25 = 0.23
Total 1.00 1.00 1.01 1.01 1.00

x) PGI-dimer
Locus Allele Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
F(A) = F(A) = F(A) = F(A) = F(A) =
2(AA)+AB+AC+AD+AE 2(AA)+AB+AC+AD+AE 2(AA)+AB+AC+AD+AE 2(AA)+AB+AC+AD+AE 2(AA)+AB+AC+AD+AE
2N 2N 2N 2N 2N
2(6)+0+0+5+0 2(1)+5+2+0+0 2(25)+0+0+0+0 2(0)+5+6+0+0 2(2)+0+0+0+1
1 =
2(25)
=
2(22)
=
2(25)
=
2(21)
=
2(25)
17 9 50 11 5
= = = = =
50 44 50 42 50
= 0.34 = 0.20 = 1.00 = 0.26 = 0.10
F(B) = F(B) = F(B) = F(B) = F(B) =
2(BB)+AB+BC+BD+BE 2(BB)+AB+BC+BD+BE 2(BB)+AB+BC+BD+BE 2(BB)+AB+BC+BD+BE 2(BB)+AB+BC+BD+BE
2N 2N 2N 2N 2N
2(2)+0+0+0+2 2(1)+5+0+2+0 2(0)+0+0+0+0 2(0)+5+0+2+3 2(3)+0+4+0+0
2 =
2(25)
=
2(22)
=
2(25)
=
2(21)
=
2(25)
Pgi – L1 =
6
=
9
=
0
=
10
=
10
50 44 50 42 50
= 0.12 = 0.20 = 0.00 = 0.24 = 0.20
F(C) = F(C) = F(C) = F(C) = F(C) =
2(CC)+AC+BC+CD+CE 2(CC)+AC+BC+CD+CE 2(CC)+AC+BC+CD+CE 2(CC)+AC+BC+CD+CE 2(CC)+AC+BC+CD+CE
2N 2N 2N 2N 2N
2(3)+0+0+3+0 2(2)+2+0+0+3 2(0)+0+0+0+0 2(2)+6+0+0+0 2(4)+0+4+2+0
3 =
2(25)
=
2(22)
=
2(25)
=
2(21)
=
2(25)
9 9 0 10 14
= = = = =
50 44 50 42 50
= 0.18 = 0.20 = 0.00 = 0.24 = 0.28
F(D) = F(D) = F(D) = F(D) = F(D) =
4 2(DD)+AD+BD+CD+DE 2(DD)+AD+BD+CD+DE 2(DD)+AD+BD+CD+DE 2(DD)+AD+BD+CD+DE 2(DD)+AD+BD+CD+DE
2N 2N 2N 2N 2N
 2(2)+5+0+3+0 2(3)+0+2+0+0 2(0)+0+0+0+0 2(3)+0+2+0+0 2(1)+0+0+2+3
= = = = =
2(25) 2(22) 2(25) 2(21) 2(25)
12 8 0 8 7
= = = = =
50 44 50 42 50
= 0.24 = 0.18 = 0.00 = 0.19 = 0.14
F(E) = F(E) = F(E) = F(E) = F(E) =
2(EE)+AE+BE+CE+DE 2(EE)+AE+BE+CE+DE 2(EE)+AE+BE+CE+DE 2(EE)+AE+BE+CE+DE 2(EE)+AE+BE+CE+DE
2N 2N 2N 2N 2N
2(2)+0+2+0+0 2(3)+0+0+3+0 2(0)+0+0+0+0 2(0)+0+3+0+0 2(5)+1+0+0+3
5 =
2(25)
=
2(22)
=
2(25)
=
2(21)
=
2(25)
6 9 0 3 14
= = = = =
50 44 50 42 50
= 0.12 = 0.20 = 0.00 = 0.07 = 0.28
Total 1.00 0.98 1.00 1.00 1.00
F(A) F(A) F(A) F(A) F(A)
= 2(AA) / 2N = 2(AA) / 2N = 2(AA) / 2N = 2(AA) / 2N = 2(AA) / 2N
=2(6+2+3+2+2+0+ =2(1+1+2+3+3+5+ =2(25+0+0+0+0+0 =2(0+0+2+3+0+5+ =2(2+3+4+1+5+0+
Pgi – L2 1 0+5+0+0+0+2+3+0 2+0+0+0+2+0+0+3 +0+0+0+0+0+0+0+ 6+0+0+0+2+3+0+0 0+0+1+4+0+0+2+0
+0) / 2(25) +0) / 2(22) 0+0+0) / 2(25) +0) / 2(25) +3) / 2(25)
= 50/50 = 44/44 = 50/50 = 42/42 = 50/50
= 1.00 = 1.00 = 1.00 = 1.00 = 1.00
Total 1.00 1.00 1.00 1.00 1.00

3. Determine the observed heterozygosity per locus (Ho) and expected heterozygosity per locus (He) for
each population.
Pop 1 Pop 2 Pop 3 Pop 4 Pop 5
Locus
Ho He Ho He Ho He Ho He Ho He
d+e+f d+e+f d+e+f d+e+f d+e+f
= =1– = =1– = =1– = =1– = =1–
N N N N N
8+6+4 (0.362 + 10+5+5 (0.382 + 5+5+4 (0.402 + 5+10+5 (0.422 + 15+0+5 (0.342 +
Adh = = = = =
25 0.362 + 25 0.422 + 25 0.302 + 25 0.242 + 25 0.442 +
18 20 14 20 20
=
25
0.282) =
25
0.202) =
25
0.302) =
25
0.342) =
25
0.222)
= 0.7200 = 0.6624 = 0.8000 = 0.6392 = 0.5600 = 0.6600 = 0.8000 = 0.6504 = 0.8000 = 0.6424
d+e+f d+e+f d+e+f d+e+f d+e+f
= =1– = =1– = =1– = =1– = =1–
N N N N N
5+10+5 (0.422 + 5+5+4 (0.402 + 8+6+4 (0.362 + 10+5+5 (0.382 + 15+0+5 (0.342 +
Per = = = = =
25 0.242 + 25 0.302 + 25 0.362 + 25 0.422 + 25 0.442 +
20 14 18 20 20
=
25
0.342) =
25
0.302) =
25
0.282) =
25
0.202) =
25
0.222)
= 0.8000 = 0.6504 = 0.5600 = 0.6600 = 0.7200 = 0.6624 = 0.8000 = 0.6392 = 0.8000 = 0.6424
d+e+f d+e+f d+e+f d+e+f d+e+f
= =1– = =1– = =1– = =1– = =1–
N N N N N
5+10+5 (0.422 + 10+5+5 (0.382 + 15+0+5 (0.342 + 5+5+4 (0.402 + 8+6+4 (0.362 +
Sdh = = = = =
25 0.242 + 25 0.422 + 25 0.442 + 25 0.302 + 25 0.362 +
20 20 20 14 18
=
25
0.342) =
25
0.202) =
25
0.222) =
25
0.302) =
25
0.282)
= 0.8000 = 0.6504 = 0.8000 = 0.6392 = 0.8000 = 0.6424 = 0.5600 = 0.6600 = 0.7200 = 0.6624
=
c =1– =
c =1– =
c =1– =
c =1– =
c =1–
N N N N N
Sod 15 (0.502 + 5 (0.702 + 0 (0.202 + 5 (0.302 + 14 (0.322 +
= 0.502) = 0.302) = 0.802) = 0.702) = 0.682)
25 25 25 25 25
= 0.6000 = 0.5000 = 0.2000 = 0.4200 = 0.0000 = 0.3200 = 0.2000 = 0.4200 = 0.5600 = 0.4352
=
c =1– =
c =1– =
c =1– =
c =1– =
c =1–
N N N N N
6Pgd 0 (0.202 + 15 (0.502 + 5 (0.702 + 14 (0.322 + 5 (0.302 +
= 0.802) = 0.502) = 0.302) = 0.682) = 0.702)
25 25 25 25 25
= 0.0000 = 0.3200 = 0.6000 = 0.5000 = 0.2000 = 0.4200 = 0.5600 = 0.4352 = 0.2000 = 0.4200
=
c =1– =
c =1– =
c =1– =
c =1– =
c =1–
N N N N N
Pgm 0 (0.202 + 5 (0.302 + 14 (0.322 + 15 (0.502 + 5 (0.702 +
= 0.802) = 0.702) = 0.682) = 0.502) = 0.302)
25 25 25 25 25
= 0.0000 = 0.3200 = 0.2000 = 0.4200 = 0.5600 = 0.4352 = 0.6000 = 0.5000 = 0.2000 = 0.4200
Aat =1– =1– =1– =1– =1–
0.0000 (1.002) 0.0000 (1.002) 0.0000 (1.002) 0.0000 (1.002) 0.0000 (1.002)
L1 = 0.0000 = 0.0000 = 0.0000 = 0.0000 = 0.0000
 =
c =1– =
c =1– =
c =1– =
c =1– =
c =1–
Aat N N N N N
20 (0.482 + 25 (0.502 + 15 (0.622 + 15 (0.502 + 14 (0.482 +
= 0.522) = 0.502) = 0.382) = 0.502) = 0.522)
L2 25 25 25 25 25
= 0.8000 = 0.4992 = 1.0000 = 0.5000 = 0.6000 = 0.4712 = 0.6000 = 0.5000 = 0.5600 = 0.4992
g+h+i g+h+i g+h+i g+h+i g+h+i
= = = = =
N =1– N =1– N =1– N =1– N =1–
Dia 5+5+0 0+0+0 3+2+3 0+5+0 2+1+2
= (0.602 + = (0.802 + = (0.562 + = (0.102 + = (0.502 +
25 25 25 25 25
L1 10 0.402) 0 0.202) 8 0.442) 5 0.902) 5 0.502)
= = = = =
25 = 0.4800 25 = 0.3200 25 = 0.4928 25 = 0.1800 25 = 0.5000
= 0.4000 = 0.0000 = 0.3200 = 0.2000 = 0.2000
c+f+i c+f+i c+f+i c+f+i c+f+i
= = = = =
N =1– N =1– N =1– N =1– N =1–
Dia 5+2+0 0+0+0 0+5+3 0+0+0 0+5+2
= (0.502 + = (1.002 + = (0.522 + = (0.002 + = (0.502 +
25 25 25 25 25
L2 7 0.502) 0 0.002) 8 0.482) 0 1.002) 7 0.502)
= = = = =
25 = 0.5000 25 = 0.0000 25 = 0.4992 25 = 0.0000 25 = 0.5000
= 0.2800 = 0.0000 = 0.3200 = 0.0000 = 0.2800
e+f+g e+f+g e+f+g e+f+g e+f+g
+i+j+k
=1– +i+j+k
=1– +i+j+k
=1– +i+j+k
=1– +i+j+k
=1–
= (0.262 + = (0.802 + = (0.402 + = (0.252 + = (0.332 +
N N N N N
1+2+5 0+0+0 1+5+3 10+0+0 3+5+2
Lap +0+2+3 0.202 + +0+1+2 0.022 + +1+1+2 0.272 + +0+5+5 0.382 + +2+3+2 0.252 +
= = = = =
23 0.242 + 25 0.042 + 24 0.172 + 20 0.132 + 24 0.192 +
13 3 13 20 17
=
23
0.302) =
25
0.142) =
24
0.172) =
20
0.252) =
24
0.232)
= 0.5652 = 0.7448 = 0.1200 = 0.3384 = 0.5417 = 0.7093 = 1.0000 = 0.7137 = 0.7083 = 0.7396
= = = = =
f+g+i+j+k+ =1– f+g+i+j+k+ =1– f+g+i+j+k+ =1– f+g+i+j+k+ =1– f+g+i+j+k+ =1–
l+m+n+o+p (0.342 + l+m+n+o+p (0.202 + l+m+n+o+p (1.002 + l+m+n+o+p (0.262 + l+m+n+o+p (0.102 +
N N N N N
Pgi =
0.122 +
=
0.202 +
=
0.002 +
=
0.242 +
=
0.202 +
0+0+5+0+0+ 0.182 + 5+2+0+0+0+ 0.202 + 0+0+0+0+0+ 0.002 + 5+6+0+0+0+ 0.242 + 0+0+0+1+4+ 0.282 +
L1 0+2+3+0+0 0.242 + 2+0+0+3+0 0.182 + 0+0+0+0+0 0.002 + 2+3+0+0+0 0.192 + 0+0+2+0+3 0.142 +
25 22 25 21 25
10 0.122) 12 0.202) 0 0.002) 16 0.072) 10 0.282)
= = = = =
25 = 0.7656 22 = 0.8076 25 = 0.0000 21 = 0.7762 25 = 0.7736
= 0.4000 = 0.5455 = 0.0000 = 0.7619 = 0.4000
Pgi =1– =1– =1– =1– =1–
0.0000 (1.002) 0.0000 (1.002) 0.0000 (1.002) 0.0000 (1.002) 0.0000 (1.002)
L2 = 0.0000 = 0.0000 = 0.0000 = 0.0000 = 0.0000
Total 5.3652 6.0928 4.8255 5.2444 4.6217 5.3125 6.0819 5.4747 5.4283 6.2348
Heterozy Ho He Ho He Ho He Ho He Ho He
5.3652 6.0928 4.8255 5.2444 4.6217 5.3125 6.0819 5.4747 5.4283 6.2348
-gosity = = = = = = = = = =
13 13 13 13 13 13 13 13 13 13
per locus = 0.4127 = 0.4687 = 0.3712 = 0.4034 = 0.3555 = 0.4087 = 0.4678 = 0.4211 = 0.4176 = 0.4796

4. Determine the inbreeding coefficient (Fis) for each population.

Population Population 1 Population 2 Population 3 Population 4 Population 5
Ho Ho Ho Ho Ho
Inbreeding Fis = 1 - He Fis = 1 - He Fis = 1 - He Fis = 1 - He Fis = 1 - He

coeffiecient 0.4127 0.3712 0.3555 0.4678 0.4176

= 1 - 0.4687 = 1 - 0.4034 = 1 - 0.4087 = 1 - 0.4211 = 1 - 0.4796
(Fis)
= 0.1195 = 0.0798 = 0.1302 = -0.1109 = 0.1293
5. Explain the genetic diversity parameters estimated for this species.

In this experiment, ten enzyme systems were used to investigate genetic diversity of five population of a
diploid species. There are three parameters were used, which are observed heterozygosity per locus (Ho),
expected heterozygosity per locus (He), and inbreeding coefficient (Fis). Heterozygosity per locus is the
probability resulting in two different alleles at the given locus of genetic diversity. Observed heterozygosity
per locus (Ho) is estimated from individual genotypes directly and depends on both the amount of genetic
variation in the population and the level of inbreeding, which increases homozygosity (Ritland 1996).
Expected heterozygosity per locus (He) is estimated from an allele frequency of each population, and this is a
common statistic for assessing genetic variation within populations (Harris & DeGiorgio 2017). Inbreeding
coefficient (Fis) is the measurement used for the probability that two genes in the same locus are identical.

Genetic variability is depending on heterozygosity in a population. The higher the heterozygosity in a

population, the higher the genetic variability. Based on the results, population 4 is the highest observed
heterozygosity, which is 0.4678. This shows that population 4 has the most genetic variability based on the
number of genetic variation in that population. While population 3 has the lowest amount of an observed
heterozygosity, which is 0.3555 and this shows that population 3 has the lowest genetic variability based on
the number of genetic variation in that population. For expected heterozygosity, population 5 has the highest
amount of expected heterozygosity which is 0.4796. This shows that genetic variability of population 5 is the
highest based on the allele frequency. While the lowest expected heterozygosity is population 2, which is
0.4034 and this shows that genetic variability of population 2 is the lowest based on the allele frequency
(Chattopadhyay 2017).

Inbreeding coefficient (Fis) is the amount of genetic diversity found between populations relative to
the amount within populations (Wright 1965). This calculates the probability that two copies of a gene variant
have been inherited from an ancestor common to both the maternal and paternal. Homozygosity will be
increases by inbreeding, while inbreeding coefficient is to calculate homozygosity in a population compared
to the average of population. Based on the results, the highest inbreeding coefficient is population 3, which is
0.1302. This shows that population 3 is the most closely related to the parents and it has the highest
homozygosity among the other populations. While population 4 has the lowest value of inbreeding coefficient,
which is -0.1109. This shows that population 4 is the furthest related to the parents and it has the lowest
homozygosity among the other populations.
REFERENCES

Chattopadhyay, N.R. 2017. 5 negative aspects of breeding practice. Science Direct.

https://www.sciencedirect.com/science/article/pii/B9780128017746000055 [21 January 2023]

Harris, A.M., & DeGiorgio, M. 2017. An unbiased estimator of gene diversity with improved variance for
samples containing related and inbred individuals of any Ploidy. G3: Genes|Genomes|Genetics 7(2):
671 – 691.

Ritland, K. 1996. Estimators for pairwise relatedness and individual inbreeding coefficients. Genetical
Research 67(2): 175 – 185.

The Kennel Club. 2023. Inbreeding calculators (Cols). https://www.thekennelclub.org.uk/health-and-dog-

care/health/getting-started-with-health-testing-and-screening/inbreeding-calculators/ [21 January
2023]

Wright, S. 1965. The interpretation of population structure by F-statistics with special regard to systems of
mating. Evolution 19: 395 – 420.