Supporting Appendix 1. Details of the genotyping procedures used in the present study
Gene
SNP
identification
Consequence
Procedure
Amplification: Primer sequences
ADH1B
rs1229984
(A/G)
ADH1B
rs1041969
(C/A)
ADH1B
rs6413413
(A/T)
Thr60Ser
Amplificationrestriction
ADH1B
rs2066702
(C/T)
Arg370Cys
TaqMan
C__11941896_20
Applied Biosystems
ADH1C
rs35385902
(G/A)
Arg48His
Amplificationrestriction
TCAGCAAAACCCTTGAGAAT
ATGGTGAACCAAGGCATGT
T
ADH1C
rs283413
(G/T)
Gly78X
Taqman
C__26457440_10
Applied Biosystems
ADH1C
rs34195308
(C/T)
Pro166Ser
Amplificationsequencing
TCCTGTCTCATGCCTTTGTG
GTCGAAAATCCACAGCCAAT
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His48Arg
Amplificationrestriction
GCTGTAGGAATCTGTC
CCTTCTCCAACACTCT
Asn57Lys
Amplificationrestriction
GCTGTAGGAATCTGTC
CCTTCTCCAACACTCT
GCTGTAGGAATCTGTC
CCTTCTCCAACACTCT
PCR conditions
94ºC 30 sec
52ºC 30 sec
72ºC 30 sec
35 cycles
94ºC 30 sec
52ºC 30 sec
72ºC 30 sec
35 cycles
94ºC 30 sec
52ºC 30 sec
72ºC 30 sec
35 cycles
92ºC 15 sec
60ºC 90 sec
45 cycles
94ºC 30 sec
55ºC 30 sec
72ºC 30 sec
40 cycles
92ºC 15 sec
60ºC 90 sec
45 cycles
94ºC 30 sec
58ºC 30 sec
72ºC 30 sec
35 cycles
Allelic discrimination
Restriction with NmuCI
A: 93+20 bp
G: 113 bp
Restriction with SexAI
C: 69+44 bp
A: 113 bp
Restriction with BstEII
A. 65+48 bp
T: 113 bp
Fluorescent dyes as indicated by
the manufacturer
Restriction with MslI
G: 295 bp
T: 141+154 bp
Fluorescent dyes as indicated by
the manufacturer
Sequencing
Gene
SNP
identification
Consequence
Procedure
Amplification: Primer sequences
ADH1C
rs1693482
(C/A)
Arg272Gln
Taqman
C___2688502_10
Applied Biosystems
ADH1C
rs698 (T/G)
Ile350Phe
Taqman
C__26457410_10
Applied Biosystems
ADH1C
rs35719513
(C/A)
Pro352Thr
Amplificationsequencing
CCCAAACTTGTGGCTGACTT
AATGGAAAACCAAGGCACT
G
ALDH2
rs671
ADH2*2 (G/A)
Glu487Lys
Amplificationrestriction
CAAATTACAGGGTCAAGGG
CT
CCACACTCACAGTTTTCTCT
T
CYP2E1
CYP2E1*5
(C/T)
Increased
gene
expression
Amplificationrestriction
GTCCCTGCCACCTCACACT
CCCTCTTCCACCTTCTATG
CYP2E1
rs72559710
CYP2E1*2
(G/A)
Arg76His
Amplificationrestriction
TGACTTCTAGCCACGGGTC
TC
TTGTAGTCCAGCAGCGCTT
CC
PCR conditions
92ºC 15 sec
60ºC 90 sec
45 cycles
92ºC 15 sec
60ºC 90 sec
45 cycles
94ºC 30 sec
57ºC 30 sec
72ºC 30 sec
35 cycles
94ºC 60 sec
52ºC 3 min
72ºC 60 sec
30 cycles
94ºC 10 sec
62ºC 10 sec
72ºC 50 sec
35 cycles
94ºC 60 sec
45ºC 60 sec
72ºC 60 sec
40 cycles
The rs numbers are shown when available. The rest of SNPs are referred to as the variant alleles that contain such SNP variant.
Allelic discrimination
Fluorescent dyes as indicated by
the manufacturer
Fluorescent dyes as indicated by
the manufacturer
Sequencing
Restriction with MboII
G: 135+125 bp
A: 260 bp
Restriction with RsaI
C: 421+144 bp
T: 565 bp
Restriction with HhaI
G: 76+54 bp
A: 131 bp
Supporting Appendix 2. Performance in reaction and motor time tests in subjects with different genotypes
PARAMETER
Reaction time
at Tmax
(msec)
Basal reaction
time (msec)
Motor time at
Tmax (msec)
Basal motor
time (msec)
ADH1B 60
SER (N=8)
ADH1B 60
NONMUTATED
(N=242)
ADH1C 78 X
(N=31)
ADH1C 78
NONMUTATED
(N=219)
ADH1C 272
GLN
(N= 150)
ADH1C 272
NONMUTATED
(N=100)
ADH1C 350
PHE
(N= 158)
ADH1C 350
NONMUTATED
(N=92)
511.1 ± 78.0
521.6 ± 95.8
512.5 ± 84.9
521.3 ± 96.7
517.0 ± 92.8
527.6 ± 98.9
516.3 ± 95.3
529.7 ± 97.7
436.1 ± 55.9
463.7 ± 87.4
455.5 ± 73.9
463.9 ± 88.5
457.8 ± 86.5
470.4 ± 87.0
458.1 ± 87.5
470.8 ± 86.1
169.5 ± 22.1
175.8 ± 60.1
182.2 ± 60.2
176.9 ± 59.6
176.5 ± 56.9
177.6 ± 63.7
177.2 ± 57.1
178.1 ± 63.8
146.5 ± 26.4
166.5 ± 57.6
174.5 ± 50.1
165.4 ± 58.2
161.8 ± 53.9
171.2 ± 61.7
164.3 ± 56.6
167.4 ± 61.6
T-test indicated the absence of significant differences between carriers and non-carriers for all mutations.
Supporting Appendix 3. Performance in reaction and motor time tests according smoking and drinking habits
PARAMETER
Reaction time
at Tmax
(msec)
Basal reaction
time (msec)
Motor time at
Tmax (msec)
Basal motor
time (msec)
SMOKERS (N=54)
NON-SMOKERS
(N=196)
DRINKERS
(N=90)
NON-DRINKERS
(N=160)
522.3 ± 116.7
521.0 ± 89.9
521.7 ± 97.7
521.0 ± 95.9
461.0 ± 109.2
463.3 ± 80.9
460.1 ± 89.6
464.4 ± 86.4
173.5 ± 68.3
178.7 ± 57.7
174.8 ± 59.8
179.1 ± 59.5
171.2 ± 69.4
164.0 ± 53.5
160.7 ± 52.7
168.3 ± 59.8
T-test indicated the absence of significant differences between subgroups.