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1. 142. PC PCR P0 30 gt KOD Plus Neo PCR KDD Plus Taq 80 KOD Plus DNA EEH DNA DNA KOD Plus Neo KOD Plus Ver 2 KOD Plus Taq
3. DNA PCPH TKOD FX DNA PCHR PCR Aspe g7 u5 oOyZg g 2005 PCR DNA TKOD FX
4. 20C gt Taq DNA polymerase 50UX 1 200 BTQ 101 19 000 Blend Tadq Blend Taq 2 5U Ll 250Ux 1 x 5 1 000 BTQ 101X5 8 000 10XBuffer mM dNTPs 250UX1 x 10 2 000 BTQ 101X10 Taq DNA polymerase Blend Tad Plus 50UX 1 600 BTQ 20 1 1 000 Blend Tag Plus 2 5U ul 10XBuff mM 250UX1 x 5 1 000 BTQ 201X5 90 000 Cell Applications Inc CAI 1 E mail Lot i Es CAI www toyobo cojpbio 12 m ua aa
5. BPB Dromophenol blue PCR 6 PCR DNA Se Quick Taqe HS Dye Mix 2X 30 4C 3 DNA 50ng globin 1 3kb 3 6kb 3 9kb Quick Taq HS DyeMix Quick Tag A HS DyeMix Taq Master Mix M 1 2 3 1 2 3 globin 1 3kb Forward Primer TTAGGCCTTAGCGGGCTTAGAC Reverse Primer CCAGGATTTTTGATGGGACACG globin 3 6kb Forward Primer GGTGTTCCCTTGATGTAGCACA Reverse Primer ACATGTATTTGCATGGAAAACAACTC Ma adder PCR 3 9kb P7 P8 1 globin 1 3kb Forward Prime
6. CAI Code No CA60205a Code No CA10205a Code No CA15005f 0 1 A Ea s ia 2 Ne ww CD54 Caspase 8 VEGF D Code No CACA054 1 Code No CACA0689 Code No CACA1461 5 8 70 10 198 152 20OFF 20 9 S02 20OFF 3 0k 20k e3039 OFF 1 0k e240 30 20FF Hsp60 primer 1 RT PCR Total RNA 1 Testis Code No CA1H51 50 2 Stomach Code No CA1H20 50 RNC SDD CA CAI www toyobo co jp bio
7. 12 DINA KOD FX DNA PCR Taq PCP KOD FX EA DNA SDS DNA RNase KOD FX PCR DNA
8. PCR DNA 2 KOD FX www toyobo co jp bio Code No KOD FX 00UX1 00 0C KFX 101 35 000 KOD FX 1U mM dNTPs 200Ux1 X10 2 000 0C KFX 101X10 60 000 XPCR Buffer for KOD FX 1 7mlx93 0C KFX 1B 5 000 50 KOD FX DNA TA TArget Clone Plus KOD TA TArget Clone Plus HOT Cell Applications INC Total RNA Cell Applications INC CAI Total RNA CAI 10
9. TA TArget Clone Plus Code No TAK 201 A TA 10 A attachment Mix KOD DNA polymerase Taq DNA polymerase PCR PCR A T 10XA attachment Mix Code No TAK 301 T 60C 10min A 15min EE bE Nk 10XA attachment Mix KOD Taq DNA polymerase T Ligase TArget Clone Plus Code No TAK 201 Ligation high Ver 2 Code No LGK 201 Total RNA ORF HeLa cell Total RNA 1 g cDNA 50ng
10. 20mer Tm gt 63C Tm Nearest Neighbor method Xl 10XPCR buffer for KOD Plus Neo 5 gil 20mer 2 2mM dNTPs 5 Eh 25 mM MgSO 3 gl 22mer Tm gt 63C 10pmol gl Primer F 1 5 gl 10pmol gl Primer R 1 5 KOD Plus Neo 1 0U 1 Genomic DNA 200ng Plasmid DNA 50nd EE ota 50 Tm 63 C Tm PCR KOD Plus Neo DNA KOD Plus PCR
11. SYBR Green 1 TaqMan i THUNDERBIRDTM 1 wm SYBR qPCR Mix 100 Code No QPS 201 tn ff TF 7 1 0 3 Co J A 100 1 me no template control 1 SYBR Green I Roche Diagnostics LightCycler 1 1 PCH Actin cDNA ReverTra Ace qPCR RT Kit Code No FSQ 101 HeLa Total RNA CDNA cDNA 10 4 no template control NTC duplicate
12. kssx 5 gt 13 www toyobo co jp bio http www toyobo co jp Seihin xr lifescience tech reader contribute index html 2 000 1 919 5 Lloyd s Register Quality Assurance LRQA ISO 900 1 Restriction Endonucleases
13. Modifying NOTICE TO PURCHASER LIMITED LICENSE PCR www toyobo co jp bio QS Accreditation R 016 Enzymes Kie eI6 5 tY AS5p JAB RN JIC 4 Quality Assurance Ltd IS014001 NN JAS ort W EE 8 IS01400 1 2
14. PCR 1 Aspergillus oryzae TE buffer 50 95C 10min 5 l 82 PCR PCR 1 5 PCR PCR grade water 6 ul x PCR buffer for KUD FX 5 ul mM dNTPs 10 Ll 10pmol ul Primer 1 1 5 ll 1 Aspergillus oryzae 10pmol ul Primer 2 1 5 ul ul KOD FX 1 0U ul 1 Ul Total reaction volume 50 Ll Target ITS 1 150bp 470bp Primer 1 GTAACAAGGT T C TCCGT Primer 2 CGTTCTTCATCGATG OAD TD PCR 84C min Seac 50 TE buffer 95C 10min
15. 1 7 5 kb 1 7 5kb PCR KOD Plus Neo KOD Plus Ver 2 A KOD Flus Neo M1 2 3 4 5 1 2 3 4 51 2 3 45 1283 4 5 5 94C 2min 94C 2min 8C 10seC 98C 10sec CVCleS 68C 30sec kb cycles 74C 3Osec KD 98C 10sec i 72C 30sec kb 98C 10sec 4 1 Human globin 1 3kb 2 Human globin 2 7kb 70C 30sec kb 9 cycles 3 Human globin 3 6kb o 4 Human globin 8 5kb 8 10S6C og cycles 5 Human globin 17 5kb 68C 30sec kb M 1kb DNA Ladder 68C 7min 1min kb 1min kb 6 PCH KOD Plus Neo 00UX1 200 KOD Plus Neo 1U ul 10XBuffer for KOD Plus Neo mM dNTP 5mM MgS04 200UX1 x5 1 000 50I PCR KOD PluS 200Ux1 200 KOD 201 30 000 PCH PCR
16. R SNOTE DNA DNA PCR DNA 10 m 1cm Xx amp 2cm 5 10 m 1 5ml 5 3 1009 5 4 70C 5 10mM Tris HCI pH 8 0 300 IP Proteinase K 10mg ml 110
17. Total RNA wwtoyopocoJpbo ED SS maSh News CELL APPLICATIONS INC Santa Cruz Biotechnology 5 000 SDS PAGE buffer 132 K 90K i 55 K 43 K
18. ORHF open reading frame 30 KOD Plus Neo ORF KOD Plus Neo A B KOD Plus Nep M 1 2 3 4 5 6 7 M 1 2 3 4 5 6 7 M 1 2 3 4 5 6 7 984C min 98C 10seC 68C 30sec kb _ lso cycles 1kb DNA Ladder Human brk Tyrosine kinase 1 6kb Human rac protein kinase alpha 1 7kb Human 63kDa protein kinase 2 0kb Human c syn protooncogene 2 2kb Human FER Tyrosine kinase 2 7kb Human cell adhesion kinase beta 3 2kb Human Jak2 kinase 3 6kb 4 ORF 1 F ATGGTGTCCCGGGACCAGGCT 3 F ACTTGACAGAATGAGGTGCGCG 5 FE AGACTTCACTGCACGTTTTCTC 7 Fi GCCCGATCTGTGTAGCCGGTTT R GACCTCTGCTGGCTTCGTGTCAC R ATCCTTCACGTGTTCTGCCAAC R TGGAGGACAGAGTCCTGAAGGC R ATCTTTGCTGGCTAGCATCATG 2 F TGTCAGCTGGTGCATCAGAG 4 F GTAGAGGCGAGGTTGTTGTGCG 6 F ACTGCAATGTGCCGATCTTAGC R GCGTGGCTTCTCTCAAATGCAC R CAGAGGAGGTTCGGATTTGGGG R TGCAAAGTGGCAAGGGAAGGT
19. Template HeLa cell Total RNA 1 g cDNA 0 5 ng 24 PCHR 40 KOD Plus Neo 4 KOD Plus Neo KOD Plus Ver 2 A B KOD Plus Nep M 1 2 3 4 M 1 2 3 4 M 1 2 3 4 M 1 2 3 4 94C min 98C 10sec 40 cycles 68C 30sec kb _ 4o ey 1 Human transferrin receptor 3 5kb 2 Human Tuberous scleosis 5 3k 3 Human adaptin 5 7kb 4 Human polymerase 6 8 kb M 1kb DNA Ladder 1min kb 1min kb 5 Template Human genomic DNA 50ng globin PCRH 30 KOD Plus Neo
20. O 55 Tmin 30 cvoles 0 ii 5 jl PCR Taq PCR PCR 30 Aspergillus O7yZg 2 PCR TOYOBO SN M 1 2 3 4 5 6 7 8 9 10 11 12 1 0 kb 1 0 kb 0 5 kb 0 5 kb 0 1 kb 0 1 kb 3 A oryzae PCR 4 PCR M 100bp ladder M 100bp ladder 6 Pichia pastris 1 KOD FX 1 Aspergillus oryzae 7 Penicillium camemberti 2 A PCR 2 Aspergillus niger 8 Penicillium requeforti 3 B Long PCR 3 Aspergillus terreus 9 Magnaporthe salvini 4 Saccharomyces cerevisiae 10 Schizophyllum commune 5 Schizosaccharomyces pombe 11 Sagenomella oligospora 12 Thysanophora penicillioides PCR 8 5 3 KOD FX
21. 1mlx1 40 QPS 201T 8 500 THUNDERBIRD SYBR qPCR Mix THUNDERBIRD SYBR qPCR Mix 1 67mlxX3 200 0C QPS 201 29 000 ON 1 67mlx3 x5 1000 20C QPS 201X5 133 000 50x ROX reference dye 50 qPCR Mix QPS 10 1X5 QPS 201X5 QPS 10 1 QPS 201 5 TaqdMan Roche Molecular Systems Inc SYBR Molecular Probes Inc PCR cDNA ReverTra Ace qPCR RT Kit Code No FSQ 101 THUNDERBIRD THUNDERBIRD Probe qPCR Mix 200 Probe qPCR RT Set ReverTra Ace qPCR RT Kit 200 OC TSPD Sa191 MM THUNDERBIRDW THUNDERBIRD SYBR qPCR Mix 200 SYBR qPCR RT Set HeverTra Ace qPCR RT Kit 200 MRNA MS THUNDERBIRD qPCR Mix 29 000 27 300 ReverTra Ace qPCR RT Kit 38 000 35 700
22. UP 1 BI CE 2 200UX1 200 6 KOD 21 1 32 000 PCR KOD FX 200UX1 200 KFX 101 35 000 TA KOD TArget Clone Plus 10 TAK 201 16 000 TA A KDD a 10xA attachment Mix 5ulx 1 25 TAK 301 12 000 Llgatlon high Ver 2 750glIx1 100 LGK 201 22 000 UPLOAD 2009 94 HOT DTaq Master Mix Quick Taqg HS DyeMix Quick Taq HS DyeMix PCR Taq i PO gr __ 5 XT Taq DNA polymerase
23. _ THUNDERBIRD x 7 Probe qPCR Mix 109 THUNDERBIRD 3 SYBR qPCR Mix 8 BP a A 0 8596 103 100 6 SYBR Green I GI cDNA 3 TaqMar GAPDH cDNA Roche Diagnostics LightCycler 1 1 Roche Diagnostics LightCyclere 1 1 1 slope 3 38 PCR 9896 slope 3 26 RSQ 1 000 ss RSQ 0 9956 20 5 1 00 0 90 0 80 0 70 0 60 0 50 0 40 0 30 0 20 0 10 0 00 10 50pl Log 2 1 414 4 SYBR Green I 10 5 SYBR Green 1 cDNA 95 Applied Biosystems 7900HT Applied Biosystems 7900HT 5 Fast Mode 50x ROX refer
24. 80 KOD FX Pfu DNA Polymerase A Long PCR Taq DNA Polymerase 00 25 0 50 0 75 0 100 0 125 0 150 0 Mutation frequency PCR PCR EE lt 30sec kb 60sec kD 30sec kbD KOD Plus Ver 2 KOD Plus Neo X M 1 2345M12 3 4 5 94C min 1 18sec 5sec kD 98C 2 6 36sec 10sec kb 68C X 54sec 15sec kD 108sec 30sec kKD 16sec 60sec kKD Tkb DNA Ladder 3 6 kb 0 Oo Sample Genomic DNA 50ng Target Human 8 globin 3 6kb 3 Ec DNA 24kbD UPLOAD voi 4 1 E
25. KOD Plus Neo p 1 rand Ne ggg PCR KOD Plus Neo ES HOT ITEM 4 Taq Master Mix Quick Taq HS DyeMix 20 02 LC THUNDERBIRD qPCR Mix U SERS NOTE KOD FX DNA TECHNICAL REVIEW 9 PCR KOD FX PCR HOTITEM 11 Cell Applications INC FLASHINEWS 12 eTransfected Lysate INFORMATION 13 13 _ 14 10 VOL 94 6 A PCR KOD Plus Neo G009 1 1 8 801 0 3 19 30sec kb PCR KOD DNA polymerase 3 5
26. Total Kit fetal CA882K05f 145 000 A fetal 5x105cells CA88205f 110 000 500ml CA881500 A 1set CA0gOk 324 K 1 PDI H 17 Code No SASC30935 A 293T Control Lysate Code No SASC 1 1 775 B PDI transfected 293T Lysate Code No SASC1 1 1237 lt 20C gt _ Total Kit adult Transfected Lysate 100ug 2G00l 39 000 293 Control L ASC ySate uaRSBCKOSa TOU non transfected 100ug 00Al 10760 0 000 A A CHO Control Lysate 100 g 00 20 000 adult vial ICAR88 05a 5X10 cells 93T Control Lysate SASC non transfected 100pg 00 T117752 U 000 500mlICAR821500 25 000
27. 1 SmM dNTPs 10pmol ul Primer F 5 DNA 20 10pmol ul Primer R 0 5 100ng KOD FX 1 0U ul 0 4 DNA DNA or 1 ga 1 Total 0 PCR 94C 15 min DNA GAY a A JSC 10 sec a 9 o 3USec _ 38 cyoles 68C 30sec DNA KOD FX DNA Proteinase K PCR DNA PCRH 300 bp 268 bp 100 bp M 100bp DNA Ladder 1 Prote
28. https secure toyobo co jp bio0 1 html cai cai form html UPLOAD 2009 94 13 10 www toyobo co jp bio
29. A CA090K 15 000 a http www scbt com transfected lysates html A 20 12 G009 11 13 11 15 3 G009 1 9 12 12 Taq DNA polymerase Blend TaqaW
30. 37C 3 4 70C 10 min PCR gt DNA Human 8 amp globin 68Dp Primer F GAAGAGCCAAGGGCAGGTAC Primer R CAACTTCATCCACGTTCACC Am J Pathol 154 67 75 1999 1 2 3 4 10mM Tris HCI pH 8 0 300 50 300gl Proteinase K buffer 1mm Proteinase K 37C 5 10 Proteinase K 2 3 Proteinase K 3 6 ul XPCH buffer for KOD FX 10 Proteinase K
31. ence dye Applied Biosystems Stratagene ROX ABI PRISM 7000 me Applied Biosystems 7300 Applied Biosystems 7900HT BioFlux Line Gene PP Biosystems Step One Plus M me oes le RS LightCycler 2 0 Bio Rad MJ iCycler iQ Mx3000P Stratagene Mx3005P Mx4000 Luemcyeerte0 lo E Es Taq TaqMan 1mlx1 40 QPS 101T B 500 THUNDERBIRD Probe qPCR Mix THUNDERBIRD Probe qPCR Mix 1 67mlx3 200 QPS 101 29 000 EN 1 67mlx3 x5 1000 QPS 101X5 133 000 SYBRY Green 1
32. inase K DNA 2 Proteinase K 70C10 3 DW 2gl Lane PCH KOD FX Code No 200U 1 KFX 101 35 000 200U X1 X5 KFX 101X5 140 000 200U X1 X10 KFX 101X10 260 000 KOD FX PCR http Awww toyobo co jp bio KOD FX UPLOAD 2009 94 P CR KOD FX PCR KOD FX PCR KDD DNA Polymerase PCR PCR
33. r CGCCAGGGTTTTCCCAGTCACGAC 2 7 globin 3 6kb Reverse Primer AGCGGATAACAATTTCACACAGGAAAC 3 PCR 3 9kb 0 2M DTaq Master Mix lt Quick Taq HS DyeMix 1 25mlx2 x10 1 000 _o0C DTM 101X10 1 25mlx2 100 DTM 101 9 800 50 HO 2 THUNDERBIRD qPCR Mix PCR THUNDERBIRD qPCR Mix Taq DNA polymerase PCH x PCR PCRH
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