ウシ白血病ウイルス検出用 Probe/Primer/Positive control
Contents
1. http wwwr takara bio coJp 17 CY415 Epoch Biosciences Inc RD NE2. MT a RNA 1 25 ul X 50 1 Probe Primer Mix for BLV 5 Xx 1 250 ul 2 Positive Control 2 Xx 105 copies ul 2 100 wl 3 EASY Dilution for Real Time PCR 1 ml 6 1 2 3 http wwwr takara bio coJp 2 CY415 Probe Primer Mix for BLV BLV tax tax
3. Ct CP Positive Control Ct bs C CH CFP Positive 0 Control CHECSDM CH SDM LEH 5DSDM CP YSDM SD SDSEiM C t rate TC Go std 10 NC PM PAM zi6 3788 ixio ruzxo sss smH10 FAM 3416 341 ixi si2xi0 il2xi0 sdH10 FAM 3029 3073 ixiO 9e0xi iOxio adH10 FAM 757 2755 ixi0 smixi 1937xi0 sd105 PAM sd106 FAM 20 FM FM Sample 1 Sample 2 Sample 3 FAM Sample4 FAM
4. Positive Control tax A2eo EASY Dilution for Real Time PCR DNA DNA TE tRNA rRNA 20 Il PCR Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice Real Time System Lite TP700 TP760 Ther
5. 4 NucleoSpin Blood Column Collection Tube 2 ml Buffer BQ2 2 350 11 000 Xg 3 5 1 5 ml 70 Buffer BE 50 ul 1 11 000 Xx 1 1 Proteinase K 740569 10 Proteinase K 6 mg Proteinase Buffer 260 ul 2 Buffer BQ2 Wash Buffer BQ2 concentrate 3 ml 12 ml 96 100 http wwwr takara bio coJp 4 CY415 V 2 1 1 o 20 2 Positive Con
6. http wwwr takara bio coJp 13 CY415 opfions UtO Auto Baseline Target BLV Threshold to 0 333035 7 Auto Baseline 5howr Threshold Baseline 5tart Well Tarnet Baseline End Well Tar Bt 4 4 I optioms Auto Auto Baseline Target BLV Threshold Aulo 0 333035 EE Jute Baseine Show Threshold Baseline Start Well Target A Baseline End Well Tarnet 8 Auto Baseline Analyze Target BLV Threshold Auto 0 333035 to Baseline Show Threshold Baseline Start Well Target 4 Baseline End Well Target 4 ee BL Auto af ET EE TE Ta En mt TE Lr WCSRm rh Hm ri TE LF HOmHEE rs Fa WF LTE EE rn RE Fe em rm 1 et Tn a Te ra EET HIHail aa mm 1 TmrE m PPmmwf Ti mm mts Ta LS TPmmmhed ap EE PT FT mmm 5 nL
7. Epoch Biosciences Inc US20020155484 WO0142505 WO02099141 US10 113 445 US09 876 830 Epoch Biosciences Inc Epoch Biosciences Inc A 2 B C W Tel 077 543 6116 Fax 077 543 1977 NANOGEN http ww
8. Er Ah T3 1 http www takara bio co jp 14 CY415 Baseline well Analysis Setting Advanced Setting Baseline Setting for well well Baseline Setting to use Use Ct Setting for Target well Baseline Start Cycle End Cycle Fire eiTHUS CT Eee mm Im IN iDGTHTTETE Tn US WT ma HG EN HG mai or HH PT He Lima ry Peerlrrml nr Ere TaD har a a mim LT SigCt We and Tarnewi Eliie Str Tnf i LN Tarat E Baswls am Ets linn End Rai Sa i Use ED Ue CT ati Dafitred tor Ta PErTL HRPHTEH Edi SEC Ed bc fF Baseline SetUp Plate SetUp Defne Targets Quencher TAMRA http wwwr takara bio coJp 15
9. PCR Hot Start PCR TakaRa Ex Tag Hot Start MA RNA DNA RNase H PR RNase H RNA 1
10. PCR 2 3 3 1 2 3
11. Stage 1 Hold Reps 1 95 10 Ef mm He er CF ler dH Stage 2 2step PCR Reps 40 95 5 64 30 Sample Volume 25 ul 6 Start run Analysis Amplification Plot Analyze tax Threshoid Baseline Auto 14 Auto Baseline Amplification Plot Option Auto Option Threshoid Auto Auto Baseline Threshold Auto Auto Baseline Analyze Baseline
12. CY41 IafHa Probe Primer Positive control lt CY224 gt Cycleave PCR BLV CY224 Cycleave PCR Reaction Mix SP CY510 CY415 PCR V 4 PCR v201309Da bovine leukemia virus BLV B DNA a BLV tax PCR 2 Probe Primer Positive control PCR DNA polymerase Tli RNase H A OO ei eh CycleavePCR Reaction Mix SP CY510
13. Positive Control 3 Positive Control 1 4 PCR lt gt DNA 1 DNA 100 ng http wwwr takara bio coJp 5 CY415 V 3 Positive Control 2 Positive Control 2 X 10 copies u 3
14. 4 PCR PCR Auto PCR Manual 2 DNA DNA DNA NucleoSpin Blood QuickPure NucleoSpin Blood QuickPure NucleoSpin Blood QuickPure 1 200 ul Proteinase K 1 25 ul 1 5 ml Buffer BQ1 200 ul 10 20 70 15 96 100 200 3 NucleoSpin Blood Column Collection Tube 2 11 000 X 9 1
15. Positive Control http wwwr takara bio coJp 16 CY415 VI Cycleave PCR Reaction Mix SP CY510 NucleoSpin Blood QuickPure 740569 10 50 250 Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice 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 VII
16. n 2 n 1 Si SID Fe Omit Omit ax Positive Control Positive Control a EE mm mm http wwwr takara bio coJp 6 CY415 5 lt gt sr PIPES Eee Ct J SD CP Gt SDM lS Ct SDM
17. CY415 Positive Control Positive Control hr Pi EFD Em TL ml EE 1 wiew Flate Layaut wlew well Table y SBlgCt Wells5 With BBlect tern ml melect tm FE i Collapse A fl Sample hh Target Na Task CT CT hE uantty BLV 1 BLW NTC Undeter 2 BLW NIC Undeter 3 E1 BLW STANDARD 38 559 38 559 1 4 EX BLW STANDARD 37 000 10 5 E3 BLW STANDARD 8 00B8 33 008 100 5 E4 BLW STANDARD 715 9 715 1 000 T E5 BLW STANDARD S354 5 3654 10 000 ES BLW STANDARD 3 179 a3 179 10O OOO A5 Sample 1 BLW UNKKOWIRNI 285 740 B74 1 998 523 10 BEg Sampls 2 BLW UNKKOWWN 27 794 4 3 944 G5H 11 Sample 3 BLY UNKKOWWIKN 27 359 27 359 5 395 478 lt gt Analysis Amplification View Well Table t Ct Positive Control Ct
18. Thermal Cycler Dice Real Time System 2 PCR FAM FAM Hold Cycle 1 95 10 2 step PCR Cycle 40 95 5 64 30 LIP El LT Hgld Time frr EH Crata Collmehm tart Hun http wwwr takara bio coJp 7 CY415 4 5 UNKN STD NTC Dye 2 n 2
19. V 4 PCR 3 PCR Thermal Cycler Dice Real Time System 7500 Fast Real Time PCR System StepOnePlus RealTime PCR System Life Technologies Thermal Cycler Dice Real Time System 1 OK EEEHI lt gt Thermal Cycler Dice Real Time System Software Absolute Quantification gt
20. a Other ti to arun The FER reactinhs contail OR Check http www takara bio co jp 11 CY415 4 Setup Plate Setup Define Targets and Samples Target name Reporter FAM Quencher None Assign Targets and Samples Passive Reference ROX 7500 Fast Real Time PCR System StepOnePlus He kr EF mrmt me the mmm tt Famiman Mari EMparmment Crz4HE 120El TF UF NHme Targots nid Spample3 Aseior Fargets ard am ED en ne Cm BF ni li Th hk Fd Fr sir biti snarlinnmi i Dr Ta EhTmweefTt TE tid CE Owne Ta ed Sarmgiat Al Targels ad mlgs Te FE ff iE LE mm EE im ei am TTF HFP TFT a TE HH mL tIWhulFPmEMNIELEIEWE IE nH Wi We Ta en nm mei ie Carmrm FEF RF EE nas gt 1 http www takara bio coJp 12 CY415 5 Setup Run Method
21. 2997 2992 Sample5 FAM z2oxiwl221xo zisxe 124xio izoxe 190x10 5 M M M M M M M M M M M M FA FA FA FA FA FA FA FA FA FA FA FA http wwwr takara bio coJp 9 CY415 7500 Fast Real Time PCR System Life Technologies 7500 Software for 7500 Fast Rea Time PCR System StepOnePlus Real Time PCR System Life Technologies StepOne Software for StepOne and StepOnePlus Rea Time PCR System 1 2 New Experiment Home Advanced Set up 7500 Software Advanced Set up rom Twrmptaf http wwwr takara bio coJp 10 CY415 3 Setup Experiment Properties Experiment name 7500 Fast Real Time PCR System 7500 Fast 96 Well StepOnePlus StepOnePlus Instrument 96 wells Quantitation Standard Other Standard 2 hours to complete a run 7500 Software en ie StepOnePlus Mi 96 wells
22. 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 from 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 0 NANOGEN http www takara bio co jp 18 CY415 http wwwr takara bio coJp 19 CY415 Iaf4R4 Tel 077 543 6116 Fax 077 543 1977 http www takara bio coJjp v201309Da
23. EASY Dilution for Real Time PCR Positive Control V 2 Positive Control 1 5 ul 1 Positive Control 2 Xx 10 copies ul EASY Dilution for Real Time PCR 2 1 5ml 5 2 X 10 4 copies ul 2 X 103 copies u 2 X 104 copies 2 Xx 10 copies uu 2 Xx 109 copies ul 3 1 5 ml EASY Dilution for Real Time PCR 90 ul 4 Positive Control 2 x 10 gt copies uu 10 ul 2 Xx 104 copies ul 5 2 X 104 copies uu 10 ul 2 Xx 103 copies ul 6 20 http wwwr takara bio coJp 6 CY415
24. mal Cycler Dice Real Time System Software Applied Biosystems 7500 Fast Real Time PCR System StepOnePlus RealTime PCR System Life Technologies 0 2 ml PCR 0 2 ml 8 strip tube individual Flat caps NJ600 1 8 0 2 ml Thermal Cycler Dice Real Time System CycleavePCR Reaction Mix SP CY510 1 2 Xx CycleavePCR Reaction Mix SP 625 ul 2 RNase Free dH20 500 3 ROX Reference Dye 25 ul 4 ROX Reference Dyell 25 ul DNA DNA NucleoSpin Blood QuickPure 740569 10 50 250 1 5 ml 1 000 200 20 ul 10 ul 4 http www takara bio co jp 3 CY415 IV V V 1 1
25. trol Thermal Cycler Dice Real Time System 1 2 x CycleavePCR Reaction Mix SP 12 5 ul EX Probe Primer Mix for BLV 5 Xx 5 ul 1 Xx or Positive Control or 5 ul 2 RNase Free dH20 2 5 ul Total 25 ul Life Technologies PCR 1 2 x CycleavePCR Reaction Mix SP 25MM TT Probe Primer Mix for BLV 5 Xx 5 ul 1 Xx or Positive Control or 5 ul 2 ROX Reference Dye or ROX Reference Dye II T 3 0 5 ul RNase Free dH20 2 ul Total 25 ul 1 CycleavePCR Reaction Mix SP CY510 2 100 ng 3 ABI PRISM 7000 7700 7300 Real Time PCR System StepOne StepOnePlus Real Time PCR System ROX Reference Dye 7500 Fast Real Time PCR System ROX Reference Dyell 1 0 5 ul 2 5 ul 1 1 3 3
26. w takara bio co jp 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 diagnostic 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 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 ll 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 Aportion 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
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