Cycleave®PCR O-157 (VT gene)
Contents
1. FAM ROX dH20 NaOH Solution 50 mM NaOH Tris HCl Buffer pH7 0 dH20 for Dilution VT1 Positive Control VT1 VT2 Positive Control VT2 20 C 1 3 4 8 20 http www takara bio co jp 3 CY217A2. RNase H RNA 1 VT1 VT2 FAM ROX VT1 VT2 CycleavePCR O 157 VT gene Screening Kit CY213 T
3. http wwwr takara bio coJp 16 CY217A Epoch Biosciences Inc Epoch Biosciences Inc U520020155484 WO0142505 WO02099141 US10 113 445 US09 876 830 Epoch Biosciences Inc
4. VT1 VT2 026 O111 O157 1217 1 FO LO Ame QO dippers 1 http www takara bio co jp 2 CY217A L 25 wl 50 Il 1 2 X Cycleave Reaction Mixture 2 X conc 625 ul 2 VT Primer Probe Mix FAM ROX 5 X conc 250 ul 3 dH20 1 ml 4 NaOH Solution 50mM 1 5 ml x 4 5 Tris HCI Buffer pH7 0 1M 1 ml 6 dH20 for Dilution 1ml 7 VT1 Positive Control 150 ul 30 O 8 VT2 Positive Control 150 ul 30 x 1 VT1
5. ROX FAM 1 VT1WT2 2 3 2 VT1 ROX FAM VT1 VT1 VT1WT2 VT1 4 3 3 VT2 ROX FAM VT2 VT2 NTN VT2 4 ROX at VT1 VT2 VT1 VT2 WN
6. Epoch Biosciences Inc GE A B C T Tel 077 543 6116 Fax 077 543 1977 NANOGEN http www takara bio co jp http www takara bio co jp 17 CY217A NOTICE TO PURCHASER LIMITED LICENSE L4 Quencher This product is for measurement of amplification detection for research use only in life sciences research industrial and environmental testing including food industry but excluding bio terrorism and bio warfare non human animal d
7. 2 PCR 3 4 PCR 4 4 l 2 DNA 100 ul 1 5 ml 2 0 ml lt 10 000 X g 10 50 mM NaOH Solution 85 ul 100 10 lt 1 M Tris HCI pH7 0 15 ul lt 2 000 10 000 X g 10 VT1 VT2
8. Z Z Tik Thermal Cycler Dice Real Time System Applied Biosystems 7500 Fast Real Time PCR System Life Technologies Thermal Cycler Dice Real Time System Lite ver 1 0 1 lt CycleavePCR Kit gt CycleavePCR Kit ver 1 0 Thermal Cycler Dice Real Time System Software Ver 4 0 PM M Plus Minus Assay gt Thermal Cycler Dice Real Time System 2 PCR
9. lt N gt P gt OK OUT gt Posi Nega ND PCR Error 1 TOUT 2 DNA OUT
10. http wwwr takara bio coJp 15 CY217A VII Takao T T Tanabe Y M Hong Y Shimonishi H Kurazono T Yutsudo C Sasakawa M Yoshikawa and Y Takeda 1988 Identity of molecular structure of Shiga like toxin VT1 from Esherichia coli 0157 H7 with that of Shiga toxin Microb Pathog 5 357 369 Jackson M P R J Neil A D O Brien R K Holmes and J W Newland 1987 Nucleotide sequence analysis and comparison of the structural gene for Shiga like toxin Ouand Shiga like toxin Il encoded by bacteriophages from Escherichia coli 933 FEMS Microbio Lett 44 109 114 3 Ito H A Terai H Kurokawa Y Takeda and M Nishibuchi 1990 Cloning and nucleotide sequencing of Vero toxin 2 variant genes from Escherichia coli 091 H21 isolated from a patient with the haemolytic uremic syndrome Microb Pathog 8 47 60 Weinstein D L M P Jackson J E Samuel R K Holmes and A D O Brien 1955 Cloning and Sequencing of a Shiga like toxin typell variant from a Escherichia coli strain responsible for edema disease of swine Bacterio 170 4223 4230 N aS VII IX Cycleave PCR O 157 VT1 VT2 Typing Kit CY222 Cycleave PCR Salmonella Detection Kit Ver 2 0 CY205 Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dic
11. Hold Cycle 1 95 10 3 step PCR Cycle 45 95 5 55 10 72C 20 3 Ios http www takara bio co jp 7 CY217A 4 Omit O BI S 9 TTE TEP ANZ NZ N _ XTX Xx ZN ROX 2 5 1 2 FAM IC ROX Blt Auto http www takara bio co jp 8 CY217A 2 NC PC NC
12. N gt FAM ROX FAM ROX IC i i as ie or may PS Ya E T I EAE EE A II AAAA AEA AEA ET d vine PC P gt FAM ROX FAM ROX IC wo sa M is i pa m x Nt FN FT i TA AAA AAS URE ISSA TAAL 3 gt http www takara bio co jp 9 CY217A 6 Io nlmiololg gt gt mura FN Femme
13. gt PCR ROX FAM Primer Probe Mix DNA 3 ND http www takara bio coJp 10 CY217A Applied Biosystems 7500 Fast Real Time PCR System Life Technologies Hold CT Cycle 1 95 C 10 snide Goi 3 step PCR Cycle 45 1 Advanced Setup New E
14. DNA 3 1 100 PCR PCR Hot Start PCR ARS TaKaRa Ex Tag HS RNA DNA RNase H
15. EGLO PCR Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice Real Time System Lite TP700 TP760 Thermal Cycler Dice Real Time System Software Applied Biosystems 7500 Fast Real Time PCR System Life Technologies 95 200 ul 20uk 10ul IV 1 PCR 2 3 4
16. Threshold Auto Auto Baseline Show Threshold Baseline Ampitication Pat ooo MOND 000301 0 000001 x a Cle Sampie 2 Ssmple 1 Auto Basalina ow 7 Teesnon 7 Basome Bart wonm Tapoa Baseine End vroh M Target A 2 Analysis Setting Analysis Settings T Analysis Settings http www takara bio co jp 12 CY217A 3 Analysis Setting Select a Target VT Target Ct Settings for Target Name T Threshold Baseline Apply Analysis Settings Threshold Amplification Plot Log Baseline Amplification Plot CT Sattings Fag Satings Adans Sets oven me comet ever gs tor TTP ENE EED TIA EAA TE uda rt tete Une Cutt St gu Ben change tee rotaga Pal se piesa Oe Ci Sertan Deft CY setines are wc to SIDJM Pue CY ler laroet sihen austors settings To cet ihe tardnes det
17. 1 5 ul x http www takara bio co jp 5 CY217A V 2 1 VT1 VI VT1 VT2 1 1 a 20 ul 1 5 ul 3 VT1 VT2 1
18. 2 X Cycleave Reaction Mixture 12 5 ul 1X VT Primer Probe Mix 5 X conc 5 ul 1X or or 5 ul dH20 2 5 ul Total 25 ul x DNA 2 3 2 3 PCR 1 http www takara bio co jp 6 CY217A V 3 PCR 3 PCR
19. 3 1 2 3 5 PCR PCR Auto PCR Manual http wwwr takara bio coJp 4 CY217A V 1
20. 2 VT2 VT2vha VT2vhb VT2vp1 171 bp CS 1 2 3 4 8 2 X Cycleave Reaction Mixture PCR DNA PCR PCR VT1 VT2 VT Primer Probe Mix
21. a gt eee TH VIN AS Sava 3 x1 VT1 VT2 x2 3 PCR 4 VT1 VT1 VT1 5 VT2 VT2 VT2 6
22. TaiDetar Deiner mmei AJTO Peine Sa Cm ALTO Det ino Ere Oniy AUTO EEO Gamage Batect a Ta Ca wewen ha VT apet Or bonge w Ver 1 Vee Donut beangs 1000000 dm 100000 Threshold 3000 000593 Sesser Stem a W000 rte pent ep enge ererig eal es w o Baseline 01 oor 0001 1 0 0001 1 6 00001 9 000008 2 a C u G n a gt amp 2 u a u Cycle neem i er Options Target VT M Thweshold Auto 3000 C Auto Baseline Show 7 Theeshola Baseline Start weim Targeta Baseline Ena Well m Target 4 http wwwr takara bio coJp 13 CY217A 10 View Well Table Applied Biosystems 7500 Real Time PCR System StepOnePlus Real Time PCR System Life Technologies StepOnePlus Real Time PCR System ROX target ROX IC FAM ROX target http www takara bio co jp 14 CY217A VI 1
23. e Real Time System Lite TP700 TP760 96well Hi Plate for Real Time NJ400 Sealing Film for Real Time NJ500 Plate Sealing Pads 9090 48 well snap plate NJ700 Flat cap for snap plate NJ720 0 2 ml Hi 8 Tube NJ300 0 2 ml Hi 8 Flat Cap NJ302 0 2 ml 8 strip tube individual Flat Caps NJ600 Cycleave TaKaRa Ex Taq Thermal Cycler Dice
24. iagnostic testing forensic testing and providing services to third parties who do not use the services for the purpose of a providing patient management or care and in which the results of such services are not included in patient records and b providing bio terrorism or bio warfare testing and specifically excluding without limitation I any human clinical therapeutic or diagnostic uses and animal clinical and therapeutic uses including any use of the results of any testing performed with any product for patient management or care or the use of the results of the services for patient management or care and II any research or services where the results of any test or assay are used for patient management care or otherwise in making therapeutic or treatment decisions for a patient A portion of this product is subject to proprietary rights of Epoch Biosciences Inc and are made and sold under license from Epoch Biosciences Inc under the patents and patent applications US20020155484 WO0142505 WO02099141 US10 113 445 US09 876 830 and corresponding patents issued in other countries There is no implied license for commercial use with respect to this product A license must be obtained directly from Epoch Biosciences Inc with respect to any proposed commercial use of this product and commercial use includes but is not limited to A the sale lease license or other transfer of this product or any material derived or produced fro
25. m it B the sale lease license or other grant of rights to use this product or any material derived or produced from it and C the sale lease license or other transfer of kits which include this product MY NANOGEN M40 Thermostable RNase H This product is covered by the claims of U S Patent No 7 422 888 and its foreign counterpart patent claims M46 PCR Cycleave This product is covered by the claims of Japanese Patent No 4022522 M57 LA Technology This product is covered by the claims 6 16 outside the U S corresponding to the expired U S Patent No 5 436 149 http www takara bio co jp 18 CY217A http wwwr takara bio coJp 19 CY217A TafdR4 Tel 077 543 6116 Fax 077 543 1977 http www takara bio co jp SII AA RNS AL v201312Da
26. waa e CY217A IdfaR4 Cycleave PCR 0 157 VT gene Screening Kit Ver 2 0 026 O111 0157 1217 1 v201312Da 0157 H7 EHEC EHEC EHEC 0O157 H7 EHEC VT1 VT2 PCR PCR DNA DNA
27. xperiment 2 Experiment Properties Quantification Standard Curve TaqMan Re agents Other Other Include Melt Curve 3 Plate Setup Define Target Target Name VT1VT2 Reporter FAM Quench er none 4 Plate Setup Define Target Target Name IC Reporter ROX Quencher none 5 Define Samples NC PC 6 4 5 Plate Layout Passive Reference none 7 Instrument 5 5 95 5 55 10 i 72 C 25 nE 8 Start http www takara bio coJp 11 CY217A 9 Analysis HA Amplification Plot Threshold Baseline Manual 1 Amplification Plot Options Target VT Target
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