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(A) Target Inserts on clones Inserts on clones (B) (B) Dual- Dual -Phase PCR Yield Phase PCR Yield A A

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(1)

PDA-PDA-MS/UniQ = Minimum Set Primers and Unique Probes Design AlgorithmMS/UniQ = Minimum Set Primers and Unique Probes Design Algorithms by Huang s by Huang et al.et al.

Dual-Dual-Phase PCR Protocol: Phase PCR Protocol:

a)a) Touch-Touch-Down PhaseDown Phase: on original templateson original templates 36.0~ 33.0oC down 0.3oC each cycle

b)b) Steady-Steady-State PhaseState Phase: on PCR yield templateson PCR yield templates 55.0oC x30 cycles

(B)(B) PCRPCR

(A)(A) M13R/F

M13R/F Virus

Virus No.No.

592592 0707

0303 TuMV 1757

TuMV 2 Poty

2

982982 1111

0303 ZaMV 1600

ZaMV 5 Poty

5

Potex Carmo Tombus Tombus Poty Poty Poty

Genus Genus

BaMVBaMV HCRSV HCRSV TBSVTBSV CNVCNV ZaMMV ZaMMV PVAPVA DsMVDsMV

Clone Clone

856856 1010

0909 7 4776

7

789789 1111

0303 3 1682

3

99 88 66 44 11 LaneLane

0404 0808 1010 0505 0707

16131613 0101

6366

12081208 0404

3910

856856 0909

4701

916916 0202

1845

414414 0606

1826

SizeSize Primers

Primers SeQSeQ

LMu LMu - - Primer Pair PCR: Target Primer Pair PCR: Target DNAs DNAs & Yield & Yield

PCR- PCR -rx rx on M13 Clones: on M13 Clones:

(A) Target

(A) Target Inserts on clones Inserts on clones (B) (B) Dual- Dual -Phase PCR Yield Phase PCR Yield A A

B B

(2)

PDA-PDA-MS/UniQ = Minimum Set Primers and Unique Probes Design AlgorithmMS/UniQ = Minimum Set Primers and Unique Probes Design Algorithms by Huang s by Huang et al.et al.

PCR PCR -rx - rx on Respective DNA: on Respective DNA:

1) 2 primers pair 1) 2 primers pair

2) 2) All 11 MinSet All 11 MinSet primers primers

LMu LMu - - Primer Pair/MS: PCR & DIG Primer Pair/MS: PCR & DIG - - Labeling Labeling

(A,B) (A,B) PCRPCR

M13R/F M13R/F Virus

Virus No.No.

832832 0505

0202 ZaMMV

ZaMMV 4 Poty

4

Potex Carmo Tombus Tombus Poty Poty Poty Poty

Genus Genus

856856 1010

0909 TBSVTBSV

77

592592 0707

0303 TuMVTuMV

22

99 88 66 33 55 11 LaneLane

0404 0808 1010 1111 1111 0707

16131613 0101

BaMVBaMV

12081208 0404

HCRSV HCRSV

856856 0909

CNVCNV

789789 0303

PVAPVA

982982 0303

ZaMVZaMV

414414 0606

DsMVDsMV

SizeSize Primers

Primers Insert

Insert

A A

B B

Dual-Dual-Phase PCR Protocol: Phase PCR Protocol:

a)a) Touch-Touch-Down PhaseDown Phase: on original templateson original templates 36.0~ 33.0oC down 0.3oC each cycle

b)b) Steady-Steady-State PhaseState Phase: on PCR yield templateson PCR yield templates 55.0oC x30 cycles

(3)

PDA-PDA-MS/UniQ = Minimum Set Primers and Unique Probes Design AlgorithmMS/UniQ = Minimum Set Primers and Unique Probes Design Algorithms by Huang s by Huang et al.et al.

9.BaMV 9.BaMV 7.TBSV

7.TBSV 6.CNV

6.CNV

8.HCRSV 8.HCRSV 4.ZaMV

4.ZaMV 3.ZaMMV

3.ZaMMV

5.PVA 5.PVA 1.DsMV

1.DsMV 2.TuMV

2.TuMV

●● ●● ●●

●● ●● ●●

●● ●● ●●

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●● ●● ●●

●● ●● ●●

●● ●● ●●

●● ●● ●●

●● ●● ●●

(1) Array

(1) Array - - Hyb CLE: PCR Hyb CLE: PCR - - dig & Ab* dig & Ab* ap|CDP- ap|CDP -star star

(4)

PDA-PDA-MS/UniQ = Minimum Set Primers and Unique Probes Design AlgorithmMS/UniQ = Minimum Set Primers and Unique Probes Design Algorithms by Huang s by Huang et al.et al.

(2) Array

(2) Array - - Hyb Color: PCR Hyb Color: PCR - - dig & Ab* dig & Ab* ap|NBT ap|NBT

9.BaMV 9.BaMV 7.TBSV

7.TBSV 6.CNV

6.CNV

8.HCRSV 8.HCRSV 4.ZaMV

4.ZaMV 3.ZaMMV

3.ZaMMV

5.PVA 5.PVA 1.DsMV

1.DsMV 2.TuMV

2.TuMV

●● ●● ●●

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參考文獻

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