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(02)2705-5066

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10

13

19

22

23

25

26

28

30

32

35

DNA

39

DNA

41

50

50

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50
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61

61

63

65

67

68

68

68

71

 

73

73

SNP

74

SNP

77

SNP

86

89

89

93

95

95

97

102

107

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109

111

111

116

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124

124

129

131

131

133

133

135

136

137

138

146

147

150

155

157

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158

159

164

165

166

166

167

169

171

171

172

180

180

180

183

185

G6PD

185

G6PD

187

G6PD

187

G6PD

188

G6PD

192

G6PD

194

G6PD

195

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202
G6PD
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206

206

207

217

219

219

222

223

229

230

230

231

233

233

234

235

242

247

247

249
251

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268

268

269



271

271

273

274

MALDI-TOF

277

SELDI-TOF

280

281

282

282

285

287

287

290

295

298

300

300

300

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307

DNA

307

310

311

313

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315

DNA

315

PCR

316

318

319

319

327

329

329

332

349

350

350

355

357

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370

370

381

383

383

392

401

402

403

403

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(02)2705-5066

421

Technology for molecular

diagnosis

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(02)2705-5066

0.1% DNA
DNA

1984 Kary Banks Mullis

1993

98%

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(02)2705-5066

Basic technology for

molecular diagnosis

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(02)2705-5066

ASO

DNA b-DNA

FISH

DNA

PFGE

PCR

RFLP

PCR

1.

2.

3.

SSCP

ARMS
ACRS
MS-PCR
CCM
DGGE
NASBA
TMA
LCR

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(02)2705-5066

-globin

Allele-specific Oligonucleotide

-globin

Hybridization; ASO

DNA

DNA

1-1

1979

single nucleotide polymorphisms; SNPs

SNP

ASO

sickle cell

polymerase chain reaction; PCR

anemiaantitrypsin
deficiency1, 2

Fluorescence in

situ Hybridization; FISH

ASO

FISH

PCR-

DNA

ASO

DNA

1520

sequence specic oligonucleotide

probe; SSOP

mRNA

ASO

-globin 1-1

-globin

metaphase

-globin

-globin

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ASO

(02)2705-5066

5'

5'

1.

3.

2.

5'
3'

A
T

3'
5'

5'
3'

A
T

3'
5'

5'
3'

G
C

3'
5'

5'
3'

A
T

3'
5'

5'
3'

G
C

3'
5'

5'
3'

G
C

3'
5'

PCR

PCR

1.

2.

3.

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1-1

FISH

FISH

1. l o c u s s p e c i f i c

1-2FISH

probes

10 kbp

FISH

2. c e n t r o m e r i c r e p e a t
probes

probe

DNA

DNAase

NICK
TRANSLATION

+ dCTP + dATP + dGTP

+ Dig - dUTP ( Biotin - dUTP)

antiDig
Avidin

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(02)2705-5066
1-2

1520 kb DNA

3. w h o l e c h r o m o s o m e

probes

DNA 1984

Schwartz Cantor

23

PFGE6

spectral karyotype; SKY

DNA

Saccharomyces cerevisiae

translocationdeletion

DNA

duplicationamplification

1.

CHEFContour-clamped homologous

22q13

electric field7

120

2.

retinoblastoma

DNA

RB1RB1 13

13q14 RB1

3.

PFGE

electroendosmosis; EEO

FISH

8, 9 plug

DNA

DNA

Pulse-field gel

electrophoresis; PFGE

loading well
1-3

DNA
DNA

DNA www.wunan.com.tw

(02)2705-5066

plug

A-

A+

B+

1-3PFGE

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10

DNA

PulseNet

DNA

1-412

DNA
DNA

Restriction

YAC'syeast articial chromosomes

10

Fragment Length Polymorphism;

11

RFLP

1985 Sir Alec Jeffreys

RFLP

1980 DNA

1996

13

3,000
2,000
1,500
1,200
1,000
900
800
700
600
500
400
300
200
100

10

11

3,000
2,000
1,500
1,200
1,000
900
800
700
600
500
400
300
200
100

1-4PFGE O157:H7
O157:H7 AvrII XbaI PFGE
M:

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marker110 O157:H7 11
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restriction enzyme

11

14

DNA 46 bp

base substitutionaddition

1-6

deletion sequence

A a

rearrangement

AAAa

DNA

aa

DNA

Cystic fibrosis; CF

1-5

RFLP

1-7

genetic ngerprinting

CFTRcystic fibrosis transmembrane

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1-5

12

conductance regulator

25%

CF

CFTR

CFTR

CF

25% CF 50%

AA

aa

Aa

Aa

Aa

aa

Aa

AA

1-6

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(02)2705-5066
1-7CF

13

Polymerase chain

aquaticus DNA

reaction; PCR

Taq DNA
72
C16

PCR

PCR

DNA

95
C

PCR 1984 Cetus

Kary Banks Mullis 15

Taq DNA

3'5' exonuclease

PCR DNA

pg DNA g

proofreading

PCR

10 pg

Kary Banks Mullis 1993

1 g PCR

Taq DNA

PCR

DNA

DNA

1/6,000 PCR

DNA

DNA Denaturation

DNA 1 kb

AnnealingExtension

1-8

Mg2+0.52.5

DNA PCR

mM

DNA

PCR

DNA DNA

RT-PCRreverse

DNA

transcription-PCRReal-time PCRPCR-

1-9

SSPsequence-specific primingNested

PCR

PCR 17

DNA
E. coli 37
C

DNA

PCR

95
C

Nested

Nested PCR PCR

Randall K. Saiki

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PCR PCR
Thermus
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1
100
90
80
70

C
60

DNA

50

10
1 2 345 6 7

1-8PCR

DNA

5'
3'

3'
5'

5'
DNA Denaturation

5'

3'

3'

5'
3'
Annealing

3'
3'
5'

Cycles

DNA

Extension

PCR

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1-9 PCR

15

template

PCR

DNA

nested primer
PCR 18 1-10

PCR

transcription-PCR; RT-PCR

Nested PCR

Reverse

RT-PCR PCR

DNA

DNA

RNA RNA

DNA

RNA DNA

Nested PCR PCR

RT-PCR RNA

Nested

DNA complementary DNA,

reverse transcriptase

DNA Template

PCR

PCR

PCR

PCR

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(02)2705-5066
1-10Nested PCR

16

HLA typing 20

DNA 1-11RT-PCR

PCR-SSP PCR

RNA

RNA RNA

21

RNA

DNA

DNAcDNA

RT-PCR

RNA

DNA

cDNA

17, 19

PCR

Poly-

PCR Taq DNA

merase chain reaction-sequence-specific

3'5' exonuclease

primers; PCR-SSP

PCR-SSP

sequence-specific primers PCR

SSP

template

Poly(A)
mRNA

3'

5'

AAAAA
TTT TT

Oligo(dT)

Reverse Transcriptase

AAAAA
T TT TT

mRNA
cDNA

cDNA

5'
3'

TTTTT

PCR

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(02)2705-5066
1-11RT-PCR

17

Sequence-Specific Primers
P3

P1

P1

P3

DR_0803

DR_0901
P4

P2

P4
P2

PCR

PCR

1-12PCR-SSP

HLA typing SSP

HLA-DR_0901

HLA DNA

P1 P3/P4

EtBr

P2 PCR

1-12 HLA typing SSP

1-12

P1 HLA-DR_0803 HLA-

HLADR_0803 PCR-SSP

DR_0901 P2

HLA-DR_0803 HLA-DR_1501

PCR-SSP

HLA-DR_0803

DNA

PCR-SSP

aerosol barrier tips

internal control

SSP DNA

P3 P4 HLA DR_0803

P3/P4 PCR

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P1/P2
pipetman
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18

Real-time

SYBR-green I

PCR
Real-time PCR PCR

DNA minor groove

quantitative real-time polymerase chain reaction;

1-13 PCR

Q-PCR PCRkinetic polymerase

SYBR-green I

chain reaction

24

PCR 22Real-time PCR PCR

Real-time PCR

TaqMan system PCR

Non-probe

23

typeprobe type

DNA

PCR

DNA

reporter

SYBR-green I

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1-13SYBR-green I

19

quencher

1-14 Real-time PCR

cycle threshold cycle; Ct

DNA

DNA PCR

1-15

DNA

Real-time PCR

DNA

threshold cycle

5'3' exonuclease

calibration curve

Taq

DNA

25

1-14

Single-Strand

Conformational Polymorphism;

SSCP

Real-time PCR PCR

SSCP 1989

DNA

PCR cycles

DNA
PCR

Taq

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1-14TaqMan

20

Fluorescent unit

Threshold

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40
PCR cycle
1-15Threshold cycle DNA

DNA PCR

DNA PCR

DNA denature

26

Orita 1989

renature

DNA

DNA DNA

non-denatured

PAGE27

DNA

band

DNA PAGE

DNA

1-16

1-16

SSCP

PCR

PCR

PCR-SSCP

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P C R (02)2705-5066

PCR
Amplification

DNA
Heat denature

Wild Type

Mutant

DNA
Renature

WT MUT

1-16SSCP

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21

. --.--
, 2009.11

ISBN 978-957-11-5842-6()
1.
415.12

98020631

5J32

(66.3)
()

(





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