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1. 60 27 14 13 48 45 50 55 27 17 10 37 35 40 35 22 13 37 25 30 34 24 10 29 15 20 34 25 9 26 10 8 4 4 50 165 106 59 36 12. 3 2 PCR Taq DNA PCR TaKaRa Taq Hot Start Version Taq DNA DNA 20 100ng PCR DNA 10 L DNA DNA RNA PCR PCR DNA
3. 165 8 1 0 105 1 0 0 58 1 0 0 0 99 49 163 164 100 0 105 105 98 39 58 59 1 13D GRY 165 Me 9 0 59 100 0 165 165 100 0 106 106 100 0 59 59
4. PCR 5000ppm 1 F PHFA Buffer 25 15C DNA 2 8 DNA 121C 20 DNA PCR 0 1
5. 3 DNA DNA 55 0 G12A 55 0 G12C 60 0 G12D 55 0 G12R 65 0 G12V 50 0 G12S 55 0 G13D 55 0 4 4 I nt FCR PCR p
6. EGFR KRAS KRAS 3 KRAS 7 KRAS G12A G12C G12D G12R G12V G12S G13D 99 6 KRAS KRAS 7 KRAS KRAS 12 13 7 G12A G12C G12D G12R G12V G12S G13D
7. DNA 1 F PHFA Buffer 25 15C DNA 2 8C 18 1 2 199
8. 2 4 3 DNA DNA 4 2 F PHFA Buffer F PHFA Buffer 25 15C PCR PCR PCR PCR PCR F PHFA DNA
9. D PCR PCR E A B C D PCR PCR PCR D PCR D
10. 2013 5 2 22400AMX01422000 2012 11 1 KRAS 84053000 OncoGuide KRAS 5 89 J 1 EGFR OL DOE NSR 2 5 dGTP ed 2 5 GTTP EGFR 3 F PHFA Buffer 3x1000pL EDTA KRAS 12 13 7 G12A G12C G12D G12R G12V
11. E PCR PCR DNA C PCR 20 PCR DNA KRAS EGFR KRAS EGFR 2
12. PCR PCR 3 3 30 25 15C 4 F PHFA 4 1 DNA DNA DNA
13. G13D DNA DNA G13D FAM DNA PCR G13D Dab DNA KRAS FEFPE 2 1X420L DNA DNA PCR NR A Solassol J et al Int J Mol Sci 12 3191 dNTP 3204 2011 FFPE FFPE KRAS
14. KRAS Kitano S et al 8th AACR JCA joint conference in Hawaii 2010 A46 p88 91 KRAS 1 3 3 1 DNA DNA DNA DNA DNA DNA DNA PCR
15. 1 DNA 12 3 EGFR 1 PCR PCR 2 DNA 1 F PHFA 250 000 1 1 DNA
16. PCR DNA DNA DNA DONA F PHFA F PHFA 1 A 20 L POCR DNA 1 DNA 12
17. T2345 6078910 1 pl wlAOOOOOOOOOOOO G12A G12C G12D G12R Gl2V G12S eisp ygeOOOOOOOOOOOO DONA PCR COFX96 PCR DONA 65 450 490nm 515 530nm F PHFA FAM 1 95C 10 95 65C FAM 30 90 90 80 70 60 50 40 30 20 10 FAM 0 5 1 0 1 5 20 25 30 35 40 45 min Microsoft Office Excel OncoGuide KRAS Mutation Detection Kit Index Sheet DNA
18. F PHFA Buffer DNA PCR CFX96 PCR CFX96 LightCycler 480 System II LightCycler 480 Software Applied Biosystems 7500 Fast Dx PCR ABI 7500 Fast System SDS Software Microsoft Office Excel 2007 100L Taq DNA TaKaRa Taq Hot Start Version 10xPCR Buffer PCR PCR 100L 3 3 1 DNA 3 1 DNA
19. DNA 3 2 PCR PCR PCR PCR 3 2 PCR 3 3 DNA PCR Taq DNA PCR TaKaRa Taq Hot Start Version Taq DNA PCR PCR 100L TaKaRa Taq Hot Start Version 1 1 1 PCR 1 10L TaKaRa Taq Hot Start Version 5U L 0 5L 10xTaKaRa Taq Buffer Mg plus 10L DNA 1 5L up to 100L 1 DNA
20. 1 2 DNA DNA 1 3 3 3 2 DNA 10 3 165 FFPE DNA KRAS
21. G12C FAM DNA G12C Dab DNA G12D DNA G12D FAM DNA FFPE G12D Dab DNA G12R DNA G12R FAM DNA G12R Dab DNA G12V DNA 108 106 G12V FAM DNA sg 59 G12V Dab DNA G12S DNA a E G12S FAM DNA 2 3 G12S Dab DNA
22. 5 2 2 1 FFPE i DNA DNA 3 Ali 2 DNA KRAS FAM DNA 1 Dab DNA 2 2 FFPE G12A DNA FFPE DNA G12A FAM DNA HE G12A Dab DNA G12C DNA HE
23. DNA PCR PCR 1 3 3 F PHFA DNA 1 _ 90C 90C x100 65C 65C 2 3 DNA KRAS
24. 20 100ng DNA 10L DNA 30 25 15C dNTP 250M dNTP PCR PCR 100 PCR TaKaRa Taq Hot Start Version 95C 3 95C 20 57C 30 50 72C 30 72C 5 4C Hold PCR PCR
25. G12S G13D KRAS KRAS 9 F PHFA 6 7 G12A G12C G12D G12R G12V 0 G12S G13D 3 1 PCR PCR PCR PCR 4
26. PCR PCR PCR PCR 3 3 F PHFA 4 F PHFA F PHFA F PHFA PCR F PHFA Buffer 2 1 F PHFA PCR F PHFA 2 F PHFA 1 PCR 95 F PHFA Buffer 75LL 170L F PHFA Buffer 25 15C PCR
27. KRAS 1 2008 11 Allegra CJ et al American Society of Clinical Oncology Provisional Chnical Opinion Testing for KRAS Gene Mutations in Patients With Metastatic Colorectal Carcinoma to Predict Response to Anti Epidermal Growth Factor Receptor Monoclonal Antibody Therapy J Clin Oncol 27 2091 2096 2009 Yoshino T et al Chnicopathological features in metastatic colorectal cancer patients with KRAS wild type Versus codon 12 and codon 13 mutant Results from a multicenter cross sectional study by the Japan Study Group of KRAS Mutation in Colorectal Cancer ASCO Gastrointestinal Cancers symposium abstr No 407 2011 540 8575 2 9 TEL 06 6941 0308 FAX 06 6941 4861 Hl alfresa 2 9 rikengenesis 5 1
28. PCR PCR 5 TE Buffer QIAamp Wash Buffer DNA DNA K PCR DNA 6 DNA
29. DNA PCR PCR PCR PCR PCR PCR 4 100V 20 20bp DNA Ladder PCR 2 Loading Buffer 1 3 PCR 54bp 60bp PCR PCR PCR
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