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1. Tail N sm 100Dp Tm 79C Wild type WT 3mm i WT primer CreER knockinD 6EH Cre Drimer 50mM NaOH 180 34 1 bp Tm 84C Vortex Aa KI KI _ 95C 10min SS 1M Tris HCI pH8 0 20 WT WT 0 a Vortex TI _ 2 000pm 10mn 6 WT KI _y 20u 0 5 2 _ 1 60 65 70 75 80 85 90 95 Temperature C 32. PCR amp cDNA SuperRiep 4028off 201 3 1 0 9 801 0 mell 0 am RMKiMfOMP6R 96 RNA PCR CDNA HEK2993 5X104cells cCDNA HEK293 Total_ RNA RHeverTra Ace qPCR RT Master Mix Code No FSQ 20 1 cDNA 40I cDNA THUNDERBIRDS SYBR qPCR Mix Code No QPS 20 1 15 House Keeping Gene PCR RNA aa Total RNA Ct
3. PCR THUNDERBIRD qPCR Mix Taq DNA polymerase PCR X PCR Au WE 4 SYBR Green 1 TaqMane PCR PCF
4. ABI PRISM 7000 iCycler iQ ABI PRISM 7700 Bio Rad MJ MiniOpticon M Applied Biosystems 7300 CFX96 Touch M Applied Applied Biosystems 7500 Mx3000P BiosystemSs Applied Biosystems 7500 Fast Mx3005P echnologies Applied Biosystems 7900HT Mx4000 Applied Biosystems StepOne M TaKaRa Thermal Cycler Dice Applied Biosystems StepOnePlus M BioFlux Line Gene Roche LightCycler 1 x 2 0 Applied Biosystems 7500 7500 Fast Agilent Technologies Mx3000P Diagnostics LightCycler Nano Mx3005P Mx4000 ROX reference dye LightCycler 480 0 1X 1 500 1X 1 50 Es Es Tad DNA polymerase SuperPrep Cell Lysis amp RT Kit for qPCR Code No SCQ 101 HeLa S83 7 5X104 5X10
5. RT 20 0 30 60 90 120 min cDNA RNA DNase I CDNA RHNase RHNA RNA DNA cDNA ReverTra Ace cDNA cDNA 1
6. Taq KOD SYBR qPCR Mix SYBR Green 1 KOD DNA Polymerase a Taq DNA Polymerase ot 70 300 bp 70 2000 bp 2 DNA GC GC 839 IGFPR cDNA 2kb Tm 3C 5C PCR PCH 20I 1 10 2 1 3 1 2cm
7. TEL 06 6348 3888 FAX 06 6348 3833 9 00 12 00 13 00 17 00 E mail tech_osaka tOoyODO jp URL http Www toyODbO CO jp Dio x A4987 K 2013 11 22 000 1
8. IGF2R GC 83 cDNA A PCH THUNDERBIRD SYBR qPCR Mix KOD SYBR qPCR Mix GC7096 pi aor Po i pik Pha a LLS LU I 5 ag 6 5 i 5 2 6 La P Th 2 1 i GC PCR ABI StepOnePlus GC 70 PCH ReverTra Ace qPCR RT kit Code No FSQ 101 HeLa total RNA cDNA cDNA 5 5 GC JJKe7 2013 vor101 7
9. 0 5 faCtOr 1 3x AGt ACt AACt factor 0 5 upg P7ep Cell Lysis amp RT Kit for dPCR THUNDERBIRD Probe qPCR Mix Code No QPS 101 A Z factor RNA Ct SuperPrep Cell Lysis amp RT Kit for qPCR HEK293 5x 30 104cellS cDNA 40I HEK2993 Total RNA 66 6ng ReverTra Ace qPCR RT Master Mix Code No FSQ 20 1 cDNA 40 cDNA THUNDERBIRD SYBR qPCR Mix Code No QPS 20 1 15 House Keeping Gene PCR Ct
10. ABI 7500 WT knock in Tm WT CreER knock in 30 WT knock in 1 3 PCRH cDNA ReverTra Ace qPCR RHT Fast Mo
11. RT 30 R2 0 983 1 10 10 15 20 25 Ct SuperPrep RNA p 1 4 1 RACE Amplification of cDNA Ends 2 3 4 PCR PCR __ _ PCR Blot 5 ELISA Enzyme linked Immuno Assay 6 2013
12. gt 26 Applied Biosystems 7000 7300 7500 7500Fast 7700 7900HT StepOne StepOnePlus Bio Rad MJ MiniOpticon CFX96 Touch iCycler iQ Roche Diagnostics LightCycler 1 x 0 Nano 480 Agilent Technologies Mx3000P 3005P 4000 TaKaRa Thermal Cycler Dice Real Time System BioFlux LineGene ROX ROX 2013 101 13 Ke7 ViroStat ViroStat 1985 Dr Douglas McAllister MONOTOPE 0MNITOPEW RABMAB 500 MONOTOPE
13. 30 3 KOD SYBR qPCR Mix KOD FX Neo 10 UPLOAD 2013 VoL 101 PCHR KOD SYBR qPCR Mix CpG PCR GC CpDG CDG 50 ChIP PCR
14. Oligo dT 1 A431 H sapiens epidermoid carcinoma cell line 2 C2C12 M musculus myoblast cell line 3 Caco 2 H sapiens colon adenocarcinoma cell line 4 CHO K1 C griseus ovary cell line 5 COLO205 H sapiens colon adenocarcinoma cell line 6 DLD 1 H sapiens colon adenocarcinoma cell line 7 HCT 15 H sapiens colon adenocarcinoma cell line 8 HDF H sapiens primary foreskin fibroblasts primary cell 9 HEK293 H sapiens embryonic kidney cell line 10 HeLa S3 H sapiens cervix carcinoma cell line 11 HepG2 H sapiens hepatocellular carcinoma cell line 12 Jurkat H sapiens T lymphocyte cell line T 13 K562 H sapiens myelogenous leukemia cell line 14 KUSA At1 M musculus bone marrow stromal stem cell line 15 L929 M musculus aneuploid fibrosarcoma cell line 16 _ MCF7 H sapiens breast adenocarcinoma cell line 17 Neuro2a M musculus neuroblastoma cell line
15. P00l 5 10 cells 0 5 10 0 5 PCH KOD PCH kb HPLC OPC Tm Tm Tm EXCEL EE 3
16. NA direct Realtime PCR Master Mix Code No QRT 101 QRT 201 1 Step PCHR 3 HeLa S3 96 x1 04cells well 100nM phorbol 182 myristate 13 acetate PMA 24 PMA 12 1 PBS SuperPrep Cell Lysis amp RT Kit for qPCRH cDNA cDNA THUNDERBIRD Probe qPCR Mix Code No QPS 10 1 IL 6 IL 1 actin TaqMan 1 A PCHR PCH A IL 6 IL 1 Ct actin ACt PMA AACt factor
17. KOD SYBR qPCR Mix Ct KOD SYBR qPCR Mix 5x10 g l KOD SYBR qPCR Mix A KOD KOD FX Neo DNA PCR KOD FX Neo
18. 15 RNA RNase U937 RNase 45 U937 1xX10 1xX103 1x10 1x10cells R 0 983 25 Total RNA C1 10 10 15 20 25 30 C1 Supe P ep W 5 RNA 35 30 8 ul cDNA 40 THUNDERBIRD SYBR qPCR Mix Code No QPS 20 1 p actin PCR 25 Ct 20 A 15 PCH A Total RHNA
19. SuperPrep Ct 33 SAP REE EE 31 EE Es 29 27 25 259 2 44 4 4 4 444 4 44444A4A4A4444 17 15 0 5 10 15 20 well no TIL 18 mIL G 4A BB actin A Ct 3 33 oe 27 25 23 il a WS WE Ww wwWwWWWWWw wwWwww 17 15 0 5 10 15 20 well no IL 1B MIL G 4 gactin 3 Ct Z lt IL 6 gt Ct IL 6 A Ct IL 6 actin SuperPrep 2 4 30 24 10 12 0 35 26 50 4 30 0 34 Ne 29 44 8 18 0 19 lt IL 18 gt A Ct IL 18 A Ct IL 18 8 actin a Supe Prep 28 62 7 93 0 15 a 33 99 13 87 0 38 28 00 5 80 0 47 9 33 26 11 99 0 34 9 SuperPrep 8B actin 1A B actin IL 6 IL 18 SE WW mm mw su ss sw ww ww WW WW Ww LE PMA IL 6 IL 18 EE 4 4 1 1 a 1 PMA 4 facCtOr
20. Taq DNA polymerase PCR KOD SYBR qPCR _Mix no template control NTC PCR GC PCH KDD DNA polymerase PCHR Taq DNA polymerase PCH Taq DNA polymerase PCH KOD SYBR qPCR Mix GC KDD SYBR qPCR Mix GC
21. 1 Homo sapiens telomerase RNA TR gene GC 6426 19bp KOD SYBR qPCR Mix PCR Taq PCR Taq i A PCHR B PCHR 1D 1 1 1 0 1 1 Olcopy Hf NM copy TEIETTYTTETTTETTTTT bE TTT TTTTTETETTTY ET ET tt TET CWCl Cycle Human actin CpG GC 73 131bp KOD SYBR qPCR Mix PCR Taq PCR Taq A PCR C PCR 4 Log 10 copy 6 Gyela GC
22. Taq DNA polymerase PCH GC GC CpG PCR X KOD SYBR qPCR Mix KOD DNA polymerase KOD SYBR Green 1 PCR 3 5 KOD exo DNA polymerase KOD 7096 GC PCR GC CDp 1 Homo sapiens telomerase R
23. Protein A gt 909 6 FITC RABMAB RABMAB Epitomics Inc OMNITOPE ViroStat gt 95 Biotin FITC HRP Recombinant Protein HIV AdenoViruS Aflatoxin Alphavirus Aspergillus Astrovirus Bacillus anthracis Bordetella pertussis Borrelia burgdorferi Bovine Papillomavirus Brucella abortus Campylobacter jejuni Candida albicans Canine Distemper virus Canine Heartworm Canine Parvovirus Chikungunya virus Chlamydia Cholera toxin Clostridia Cryptosporidium parvum Cytomegalovirus Enolase Dengue virus Entamoeba histolytic
24. Taq DNA polymerase PCH kb Taq DNA polymerase PCHRH KOD SYBR qPCR Mix KOD SYBR qPCR Mix 1mlX1 40 QKD 201T 9 800 KOD SYBR qPCR Mix 50xXROX reference dye 1 67mlx3 200 QKD 20 1 3 000 1 67mlX3 x5 1 000 QKD 20 1X 5 147 000 50xROX reference dye 50I KOD SYBR qPCR Mix 01JKe7 12 2013 101 Es PCR KOD SYBR qPCR Mix KOD SYBR qPCR Mix Taq KOD SYBR qPCR Mix KOD DNA Polymerase
25. 1 0I USER SINOTE DNA PCR A qPCR RNA KOD SYBR qPCR Mix DNA VIOGENE Plant Genomic DNA Extraction Miniprep System DNA rDNA ITS 105Dp F 19 Tm 53 9 R PO Tm 58 4 KOD SYBR qPCR Mix A
26. PCR amp cDNA SuperPrep Cell Lysis amp RT Kit for qPCR HOT 7 7 PCOR THUNDERBIRD qPCR Mix PCRH KOD SYBR qPCR Mix US 7S NOTE KOD SYBR qPCR Mix TECHNICAL REVIEW PCR KOD SYBR qPCR Mix CpG Q amp A PCR KOD SYBR qPCR Mix NEW RELEASE ViroStat ldeas amp Chemistry SuperPrep Cell Lysis amp RT Kit for qPCR p 1 4 Ms to PCR amp cDNA SuperPrep Cell Lysis amp RT Kit tt ie for gPCR G013 10 9 0 1 4 3 80 PCR CDNA
27. ReverTra Ace qPCR RT Master Mix Code No FSQ 201 THUNDERBIRD SYBR qPCR Mix Code No QPS 201 RHNA P0C SCQ 101 6 000 SuperPrep Cell Lysis amp RT Kit for qPCR lt Lysis Reagents gt lt RT Reagents gt 100 Lysis Solution 5 x RT Master Mix Stop Solution 5 x RT Master Mix no RT control gDNA Hemover Nuclease free Water 100 x5 RNase InhiDitor SCQ 101X5 Cell a SuperPrep A RNA 6 U937 actin 325 000 SuperPrep THUNDERBIRD Probe qPCR set 3 SuperPrep Cell Lysis amp RT Kit for qPCR SCQ 101 THUNDERBIRD Probe qPCR Mix QPS 101 SuperPrep THUNDERBIRD SYBR qPCR set 4 SuperPrep Cell Lysis amp RT Kit for qPCR SCQ 101 THUNDERBIRD SYBR qPCR Mix QPS 201 SCQ101 QPS101 SCQ101 QPS 0 1 99 750 99 750 SuperPrep Cell Lysis Kit for qPCR S Lysis Solution gDNA Remover Stop Solution RHNase Inhibitor 40l
28. 100 50 200 20I 500 3 SuperPrep Cell Lysis amp RT Kit for qPCR 100 76 000 72 450 THUNDERBIRD Probe qPCR Mix Code No QPS 101 200 29 000 27 300 4 SuperPrep Cell Lysis amp RT Kit for qPCR 100 76 000 72 450 THUNDERBIRD SYBR qPCR Mix Code No QPS 201 200 29 000 27 300 5 SuperPrep Cell Lysis amp RT Kit for qPCR Lysis Reagents 1 step PCRH TaqMan RNA direct Realtime PCR Master Mix SYBR GreenI FRNA direct SYBR Green Realtime PCR Master Mix 50 100 0 250 0 5mlX5 0 5mlX5 SCQ 0 1 39 000 QRT 101 QRT 80 1 SYBR Molecular Probe Inc TaqMan Roche Molecular Systems Inc 4 33 000 33 000 HOT PCR THUNDERBIRD qPCR Mix
29. 1 G PCR THUNDERBIRDe qPCR Mix i B dl es FY YY C A A C 1 109 A 3 J F p MW 1 04 104 IM 8 EL EE 1 SYBR Green I GI 6 TaqMan GAPDH cDNA cDNA Roche Diagnostics LightCycler 1 1 Roche Diagnostics LightCycler 1 1 Fast Mode 50x ROX reference dye Applied Biosystems Agilent Technologies ROX
30. 18 NIH 3T3 M musculus embryo fibroblast cell line 19 PC12 RR norvegicus adrenal pheochromocytoma cell line 20 MSC RR norvegicus bone marrow stromal stem cells primary cell 21 THP 1 H sapiens acute monocytic leukemia cell line 22 U937 H sapiens leukemic monocyte lymphoma cell line DNasei DNase I Stop Solution RNA PCH SGYBRS Green I TaqMan A EF j E THUNDERBIRD qPCR Mix Code No QPS 101 QPS 201 p 5 p 6
31. PCHR qPCR mix Primer F Primer R Template DDW 10 2 ul 2 ul Tul lo 24 0 KOD SYBR qPCR Mix gt 98C min 98C 10sec 60C 10sec 68C 30sec x45 60C to 99C lt A qPCR gt 95C 10min 95C 15sec 60C 1min x40 60C to 95C TaKaRa Thermal Cycler Dice Real Time System Single W JKe7 2 or101 1 A qPCR a Ei mm 13 8458 7 Cie b Fai Nm Ch Fa EE KOD SYBR qPCR Mix a rT i Fi ST wm EE FEREEPE ERE ETT FE b We 3 1 Cd m1 5X10g dl 5X10 g 4 5XX10 9 i a COT TE TO EE 5X10 g d Tnere 5X10 g dl
32. 11 6 2014 1 31 ldeas amp Chemistry 7 RECORD PROPER RANDOM NESTED WESTERN INVERSE SOLVENT LIGATION KINATION COMPETENT_DIGESTION HYDRATION FORMATION CHROMATIIN SUSPEND UPLOAD 5 2 000 www toyobo co jp bio http www toyobo co jp seihin xr lifescience tech reader duiz index html 3 530 8230 2 8 TEL 06 6348 3786 FAX 06 6348 38383 E mail order lifescience toyobo jp 141 8633 _ 10 TEL 03 6422 4819 FAX 03 642 495 1 E mail order lifescience toyobo jp
33. Mix 1mlx1 40 QPS 101T a 1 67mix3 200 QPS 101 29 000 1 67mlx3 x5 1000 QPS 101X5 133 000 THUNDERBIRD SYBR qPCR Mix 1mlx1 40 QPS 201T 8 500 THUNDERBIRD SY8HY GPCR Wi 50590 1 67mlx3 x5 1000 SuperPrep THUNDERBIRD Probe qPCR set SCQ101 SuperPrep Cell Lysis amp RT Kit for qPCR SCQ 101 20D QPS101 99 750 THUNDERBIRD Probe qPCR Mix QPS 10 1 SuperPrep THUNDERBIRD SYBR qPCR set SuperPrep Cell Lysis amp RT Kit for qPCR SCQ 101 1 Os 99 750 THUNDERBIRD SYBR qPCR Mix QPS 201 50x ROX reference dye 50 Il qPCR Mix QPS 101X5 QPS 201X5 QPS 10 1 QPS 20 1 5 TadMan Roche Molecular Systems Inc SYBR Molecular Probes Inc SuperPrep Cell Lysis amp RT Kit for qPCR 100 76 000 72 450 THUNDERBIRD Probe qPCR Mix Code No QPS 101 200 29 000 27 300 2 SuperPrep Cell Lysis amp RT Ki
34. 10 1X10 1X10 1X10cellsS cDNA 40I CDNA THUNDERBIRD Probe qPCR Mix Code No QPS 101 C D PCH actin TaqMan 35 PCR 2 NN THUNDERBIRD 30 4 Probe qPCR Mix Ct 25 20 i 15 0 1 2 3 4 5 Log Cell Number m C AD THUNDERBIRD Probe qPCR Mix C D 0 EN 1x10 cells 1x101cells 1x103cells 1x102cells 1x102cells 1x104cells 1x103cells 1x103cells 5x104cells 1x104cells 1x104cells 5 7 5x104cells Sx10 cells 5x104cells 0 a 7 5x104cells a 7 5x104cells 7 1 N Bl oo 3 1 5 Cytle THUNDERBIRD Probe qPCR
35. NA TR gene GC 64 19bp CCCGCCCGAGAGAGTGACTCTCACGAGAGCCGCGAGAGTCAGCTTGGCCAATCCGTGCGGTCGGCGGCCGCTCCCT TTATAAGCCGACTCGCCCGGCAGCGCACCGGGTTGCGGAGGGTGGGCCTGGGAGGGGTGGTGGCCATTTTTTGTCT AACCCTAACTGAGAAGGGCGTAGGCGCCGTGCTTTTGCTCCCCGCGCGCTGTTTTTCTCGCTGACTT Primer 1 CCCGCCCGAGAGAGTGAC 18bp Tm 68 6C Primer AAGTCAGCGAGAAAAACAGC 20bp Tm 61 1C Human 7 actin CpDG GC 73 131bp BCTTCCTITTGTCCCCAATCTGGGCGCGCGCCGGCGCCCCCTGGCGGCCTAAGGACTCGGCGCGCCGGAAGTGGCCA GGGCGGGGGCGACTTCGGCTCACAGCGCGCCCGGCTATTCTCGCAGCTCACCATG SST GS AGG Primer 1 GCTTCCTTTGTCCCCAATCT 20bp Tm 64 1C Primer 2 CATGGTGAGCTGCGAGAATA 20bp Tm 63 9C KOD SYBR qPCR Mix 50xROX 10pmol ul Primer 1 10pmol ul Primer 2 Human genomic DNA DW total Applied Biosystems StepOnePlus 104 0 4 0 4 1 0 4 1 Xl Yl Opnul PCR 98C 2 min 98C 10 sec 4 60C 10 sec 40 cycles 68C 30 sec Melting Data collection Taq PCR PCR MMKe7 2013 vor101 1 1
36. RD SYBR qPCR Mix KOD SYBR qPCR Mix Taq 10 0P10Z1O 100 1 KOD FX Neo 2 KOD FX 3 KOD Plus Ver 2 DNA 6 TGF 1 9 8DD a md PP a a 1 PCR ABI StepOnePlus KOD FX KOD Plus 6kb Taq DNA polymerase oOkb kbD PCHRH sy a A Ls 2 hb 4 KOD DNA polymerase Taq DNA polymerase GC 709
37. SuperPrep Cell Lysis amp RT Kit for qPCR Code SCQ 10 1 PCRH Lysis Reagents RT Reagents 96 RNA RT PCR cDNA SuperPrep Cell Lysis Kit for qPCR Code SCQ 201 Lysis Reagents 1 step RT PCR Cell Lysis Revers Transcription PCR 2 7min 25min Lysis Reagents RT PCR ka Lysis Solution a Stop Solution Lys
38. a Diphtheria toxin Enterococcus E coli Enterovirus Epstein Barr virus Feline Immunodef virus Feline Leukemia virus Flavivirus Francisella tularensis Herpes Simplex virus Giardia HIV Helicobacter pylori Human Papilloma virus Hepatitis A B C virus Influenza virus Klebsiella Legionella Leptospira Lipid A Listeria Lyme B burgdorferi M tuberculosis Marburg virus Metapneumovirus Mumps virus Mycoplasma Neisseria Norovirus Pichia pastoris Papillomavirus Plasmodium Parainfluenza virus Poliovirus Parvovirus Prion Protein Pseudomonas Rabies virus Respiratory Syncytial virus Rhinovirus Rotavirus SARS coronavirus Rubella virus Shiga toxins S cerevisiae Shigella Salmonella Staphylococcus Streptococcus Tetanus Toxic Shock toxin Treponema pallidum Trichomonas Ureaplasma parvum Varicella Zoster virus West Nile Virus Yellow Fever virus Toxoplasma gondii Vaccinia virus Yersinia pestis lt www toyobo co jp bio gt ViroStat m
39. de 50xROX reference dye ROX KOD SYBR qPCR Mix 1mlx1 40 QKD 201T 9 800 KOD SYBR qPCR Mix 50xROX reference dye 57mlx3 200 QKD 20 1 32 000 1 67mIlx3 X5 1 000 QKD 201X5 147 000 50xROX reference dye 50I KOD SYBR qPCR Mix SYBR Molecular Probes Inc PCR cDNA ReverTra Ace qPCR RT Kit FSQ 101 38 000 PCH cDNA FsQ 201 38 000 ReverTra Ace qPCR RT Master Mix PCR cDNA DNA FSQ 301 40 000 ReverTra Ace qPCR RT Master Mix with gDNA Remover 901JKe7 2013 yr101
40. is Solut A Rn UUUUUUUU gDNA Remover cDNA PCR RR Lysis Reagents Supe P ep Cell Lysis Kit for qPCR Code No SCQ 201 S PCHR 1 THUNDERBIRD Probe qPCR Mix 4 a Code QPS 101 4 Oe THUNDERBIRD SYBR qPCR Mix Code QPS 201 4 RT Reagents 5xRHT Master Mix 5xXRHT Master Mix no RT control 1 DD a 2 PCR SuperPrep SuperPrep 5 Cell Lysis Kit for qPCR Cell Lysis amp RT Kit for qPCR SuperPrep Cell Lysis amp RT Kit for qPCR Code No SCQ 201 Code No SCQ 101 Code No SCG 101 2 1 N_ RNA PBS Lysis Solution gDNA Remover 5 DNA Stop Solution
41. t for qPCR 100 76 000 72 450 THUNDERBIRD SYBR qPCR Mix Code No QPS 201 200 29 000 27 300 UPLOAD 2013 vor101 6 HOT PCR KOD SYBR qPCR Mix GC KOD SYBR qPCR Mix KOD DNA polymerase KOD SYBR Green 1 PCR 3 5 KOD exo DNA polymerase KOD PCR KOD DNA Polymerase kb NN THUNDERBI
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