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PCRM-PCR PCR MPCR Dualreal PCR PCR (M-PCR) PCR (M-PCR) Q & A PCR DNA 50 2030 DNA PCR 1 PCR (MPCR) MBI PCR 1 MPCR 1 MPCR DNA 1 PCR 510

DNA (US patent 5, 582, 989) 1 DNA ( 1) MPCR GC MBI MPCR PCR DNA

2 MBI MPCR RNA mrna mrna mrna Real-Time PCR Real-Time PCR MPCR MPCR MPCR PCR (Fig.2) MPCR PCR Table1 MBI MPCR MPCR 3 MPCR mrna MBI MPCR cdna

MPCR 3 1 Short Hand on Time MBI MPCR () 4 MBI 3 MPCR MBI MBI MPCR ( 5)MPCR MPCR

PCR DNA PCR mrna MPCR PCR

MBI PCR MPCR SNP 1. HPV MPCR MPCR MPCR (HPV) 2 40 HPV HPV in vitro (HPV1618) (HPV3133) (HPV611) MBI HPV MPCR (Fig6)MBI HPV MPCR Dr. Chudzio MBI HPV MPCR HPV

2. : MPCR MBI Linden (Factor II) G20210A MPCR Figure7 Factor V G1961A (Arg506 Gln) 1994 Bertina prothrombin (factorii) G A (20210) ASA PCR PCR factor IX PCR Figure7 DNA MBI Factor V Linden G20210A MPCR Kaiser Permanent North Regional Laboratory Blood Bank

(DMD/BMD) X DMD/BMD DMD 3 DMD 70 DMD 60% Beggs Chamberlain MBI DMD/BMD MPCR 98% MPCR 1. /Toll-Like Receptors

CD14 TLR-4 (Toll-like receptor-4) TLR MBI MPCR (MP-70190) Toll-like receptor (Cao) CD14 TLR-4 (0.03mM) (1.2mM) TLR-4 CD14 631bp 203bp GAPDH 8 MPCR semi-quantitate 2. Dr. Ruediger Salowsky (Agilent Technologies Deutschland GmbH) RT-MPCR microfluidic amplicon end-point 11 Dr. Salowsky ( 9) RT-MPCR ( 9 Dr. Ruediger Salowsky Rainer Wittig ) MBI MPCR Agilent 2100 Bioanalyzer Agilent 2100 Bioanalyzer

MPCR 2 PCR PCR MPCR 10 cdna MPRCR 2 1) MPCR PCR 2) 5ng total RNA (10 ) intensity 50ng (25 ) 1/10 (28 ) MPCR

MPCR MPCR 3 MPCR PCR PCR PCR MPCR MPCR MPCR PCR ( 11) MPCR PCR PCR MPCR MPCR PCR PCR

PCR Dualreal PCR MPCR Real PCRDual quantitative PCR Real Time PCR MPCR Real-Time PCR MBI Dual Quantitative PCR MBI Dual Quantitative PCR

Quantitative PCR Dual Quantitative PCR PCR 13 A 5 cdna Real-Time PCR Dual Quantitative PCR B B MBI Real-Time PCR MBI Dual quantitative PCR MBI GAPDH (G) 18S (S)T-Transferin Receptor (T)P-Phospholipase A2 (P) L32 (L) Dual quantitative PCR Real-Time PCR

Dualreal PCR MPCR Real PCRDual quantitative PCR Real Time PCR MPCR Real-Time PCR MBI Dual Quantitative PCR MBI Dual Quantitative PCR Quantitative PCR Dual Quantitative PCR PCR 13 A 5 cdna Real-Time PCR Dual Quantitative PCR B B MBI Real-Time PCR MBI Dual quantitative PCR MBI GAPDH (G) 18S (S)T-Transferin Receptor (T)P-Phospholipase A2 (P) L32 (L) Dual quantitative PCR Real-Time PCR

PCR (M-PCR) PCR MPCR I. PCR 1. 2. 3. () II. PCR cdna 1. PCR cdna 2. PCR cdna 3. cdna PCR

III. 1. (2) PCR cdna PCR DNA PCR /cdna Taq polymerase Taq dntp dntp 20 Note / V "hot-start" 2.

(2) (5uL ) Taq polymerase V "hot-start" 3. DNA PCR PCR DNA PCR DNA 4. Taq polymerase () Perkin-Elmer AmpliTaq (AmpliTaq Gold Perkin-Elmer )

Gibco-BRL ( 2 ) BMB 5. 2 PCR 3 PCR PCR 94-96 2 2 cdna 6. Perkin-Elme III. cdna cdna PCR cdna PCR 1. DNA DNA dntp. Taq polymerase

cdna 2. cdna DNA RNA/DNA DNA dutp/udg DNA RNA RNase-free Dnase cdna MPCR cdna 3. cdna PCR PCR DNA ladder PCR PCR DNA ladder cdna DNA RNA RNase-free Dnase

4. PCR PCR DNA RT cdna PCR 5. cdna cdna mrna mrna cdna cdna RNA cdna RNA cdna RNA Human/mouse/rat/ DNA RNA Human/mouse/rat/ cdna RNA. Hot start Hot start PCR Hot start 60 Hot start Hot start 60

AmpliTaq Taq polymerase Ampliwax Perkin-Elmer 3 polymerase temperature-sensitive Clontech Taq antibody PCR (M-PCR) Q & A PCR MPCR Q1. MPCR PCR A1. PCR 1 MPCR 1 PCR PCR DNA Q2. MPCR PCR A2. MPCR Taq polymerase (Additive) MPCR DNA MPCR PCR Q3. MPCR DNA A3. MPCR PCR

Q4. MPCR A4. mrna Q5. MPCR A5. Q6. 2 PCR 3 PCR A6. PCR 94/1min.40-60/2-5min. 70/2-5min. 3 PCR 2 PCR 94/1min. / 60-70/2-5min.Taq polymerase 722 PCR 60-70 PCR 2 PCR MPCR 100 25-40 Q7. MBI PCR PCR A7. RT-PCR cdna RT-PCR MBI Q8. MPCR ampli-taq Gold A8. Taq Gold Q9. MBI

A9. MBI MPCR MPCR PCR Q10. MBI? A10. Tm 2-5 Taq polymerase Q11. A11. 2 Taq polymerase PCR Q12.? A12. 2

Q13. MPCR RNA A13. 10 6-10 7 (RNA) 100uL - cdna RNA 5uL 50uL 1:20 PCR cdna 1uL - 1:50 10 6-10 7 RNA 1:1000 10 6-10 7 total RNA Maxim Multiplex PCR Kit cdna