Cycleave® RT-PCR Giardia (18S rRNA)
Contents
1. Epoch Biosciences Inc U520020155484 WO0142505 WO02099141 US10 113 445 US09 876 830 Epoch Biosciences Inc Epoch Biosciences Inc A2. PCR RNA RNase RNA RNase RNase OFF 9037 RNA RNA http wwwr takara bio coJp 7 CY231 Vi
3. 2 3 Thermal Cycler 37 15 85 5 4 C 20 http wwwr takara bio coJp 8 CY231 VII 2 PCR 1 18S rRNA DNA 1 1
4. 1 1 0 2 ml 23 l 1 RNase Free dH2O 2 ul 1 2 x Cycleave Reaction Mixture Giardia 12 5 ul TX Giardia Primer Probe Mix FAM ROX 5 0 ul 1 RNase Free dH2O 5 5 ul 2 2 ul 2 RNase Free dH2O Total 25 ul 1 2 3
5. 4 PCR PCR Auto PCR Manual http wwwr takara bio coJp 5 CY231 VI 2 A EE 19 3 30 033006 24 3 2
6. V 1 PCR 2 3 3 1 2 3
7. http www takara bio cojp 25 CY231 IX IX 1 Giardia lamblia Assemblage A Giardia Primer Probe Mix Giardia lamblia Assemblage B Assemblage C Assemblage D Assemblage E Giardia psittaci Giardia microti IX 2 Cryptosporidium parvum X 1 Kwannan N et al 2007 Journal of Microbiology 45 2 581 583 2 Richad M Hopkins et al 1997 J Parasitol 83 1 44 51 XI Cycleave RT PCR Cryptosporidium 18S rRNA Detection Kit CY230 Cycleave PCR Legionella 16S rRNA Detection Kit CY240 CY2405S Cycleave PCR Legionella 5S rRNA Detection Kit CY210 Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice Real Time System Lite TP700 TP760 0 2 ml 8 strip tube individual Flat Caps NJ600 96 well Hi Plate for Real Time NJ400 Sealing Film fo
8. tRNA rRNA ll 20 I 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 200 ul 20 ul 10 ul VI http wwwr takara bio coJp 4 CY231 IV 1
9. RNase H RNA KI 1 Hot Start PCR ERO TaKaRa Ex Tag HS 18S rRNA FAM DNA DNA ROX 1 FAM DNA ROX PCR
10. B C AA NANOGENM Tel 077 543 6116 Fax 077 543 1977 http www takara bio coJp 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 I any human clinical therapeutic or diagnostic uses and animal clinical and therapeutic uses including any use of the results o
11. RNase Free dH20 PCR http www takara bio coJp 3 CY231 Lysis Buffer VI 40 mM NaCl 0 2 Triton X 100 4 mM DTT TE Buffer Proteinase K VI dH2O for Dilution 4 dH20 for Dilution VI Proteinase K Giardia Positive Control DNA EASY Dilution for Real Time PCR DNA DNA TE
12. 1 E 0 01 Tween 80 0 1M 50 ul Vortex 10 10 2 10 Vortex 3 10 4 1 5 ml 5 0 01 Tween 80 0 1 M 50 ul Vortex 10 6 10 1 4 7 100 ul 1M 10 8 TE buffer 10 mM Tris HCI pH7 6 1 mM EDTA 200 ul 12 000 rpm 3 260 u 50 u 20
13. 2 3 121 20 2 5
14. 0 2 ml 23 ul 1 RNase Free dH2O 2 ul 1 2 x Cycleave Reaction Mixture Giardia 12 5 ul 1x Giardia Primer Probe Mix FAM ROX 5 0 ul 1X RNase Free dH20 5 5 ul Giardia Positive Control 2 pul 2 RNase Free dH2O Total 25 ul 1 2 2 Giardia Positive Contro 2 PCR PCR 10 2 3 Giardia Positive Control 2 ul
15. FAM ROX C _ 9 t n E j i KY PE u frr EE 2 taan tirai STD lt 5 gt FAM 7 ROX FAM ROX IC s i H 10 Cj 9 404 0 0 4 3 o t2 14 l6 tt 26 22 24 20 30 2012 34 30 3 40 42 4l av 5O 9ra 4534 http www takara bio coJp 22 CY231 3 gt 4 wy JS 2 77 E4 Omit 1 0Omit 2
16. 3 4 5 http www takara bio coJp 21 CY231 5 1 FAM ROX DNA IC Bif ld4 Auto 2 NC STD NC N gt FAM ROX
17. http www takara bio coJp 20 CY231 8 Eit 4 Omit RHES Multiplex Ea mma ERE danin ee Fan syama TH f TF s 1 ZT x 2 E KO 2 400 402 129 H VN Na la 3 an n deri tE TER r pn UNKN Unknown STD Standard NTC No Template Control 1 2 3 4 1 2
18. 18S l a Es 2 3 PCR 4 18S rRNA HZD Giardia Positive Control 5 http wwwr takara bio coJp 17 CY231 VIII Giardia Positive Control gt 8 Glardia Positive Control
19. 210 Ul 100 u 50 ul http wwwr takara bio coJp 6 CY231 1 50 80 37 5 2 Lysis Buffer 149 ul amp dH20 for Dilution 12 4 Proteinase K 1 2 1 ul 10 60 30 1 2 Proteinase K G 2 4 75 10 5 95 5 80 PCR
20. http www takara bio coJp 23 CY231 UNKN Sample 36 22 UNKN Sample 36 89 UNKN Sampla 2581 UNKN Sample 2 23 NTC NC NTC NC STO 36 96 STO 37 21 STD 35 03 STO 35 22 Tb 32 30 STO 3205 BTH 2316 ETD 29 13 STD 25 85 STD 25 30 sD 2241 STO 2253 39 58 36 56 26 02 2 2 37 09 37 09 3513 35 13 32 18 32 18 2915 2915 25 88 2588 2247 2247 CCP 90 P 1980 7 RE 1 000Ex000 1 000 000 1 000E 011 1 00gs001 1 000E 002 1 000Ex002 1 000E 003 1 000E 003 1 000E 004 1 000E 004 1 0008 005 1 000E 005 3 229E 000 1 919E 000 1041E 004 7 5 6E 3 1 818E 000 1497E 000 8 128E 000 2 14E 000 6 753Ex001 211E 001 7 735E G02 7 917E 603 1 099E 004 3 709E 003 1457E 005 1 327E O05 2 5248 000 8964E 093 645 063 1 597E 000 105764000 7571E 090 7676 000 7 487E 001 7407E 001 7 926E 002 7826E 002 8 001E 003 9 901E D03 1 392E 095 1 392E lt 005
21. 2 Giardia Positive Control 3 EASY Dilution for Real Time PCR 2 6 1 2 ul n 2 1 X 105 copies ul Giardia Positive Control 1 X 104 copies ul Giardia Positive Control ul EASY Dilution 45 ul 1 X 103 copies ul 2 1 X 104 copies ul ul EASY Dilution 45 ul 1 x 102 copies ul 3 1 X 103 copies u LA 5 ul EASY Dilution 45 ul 10 copies ul 4 D 1 X 102 copies ul ul EASY Dilution 45 ul 1 copy ul 5 10 copies ul ul EASY Dilution 45 ul ON ne RD 3 3 2 0 2 ml PCR 1
22. Giardia Assemblage A E G a 7a Positive Control 0302 2 4 24 3 2 PCR O Ore AO C 1 http www takara bio coJp 2 CY231 Il 25 ul 50 1 5 x PrimeScript RT Master Mix 5 X conc 100 ul O 2 2 X Cycleave Reaction Mixture Giardia 2 X conc 625 ul 3 Giardia Primer Probe Mix FAM ROX 1 5 X conc 250 ul 4 RNase Free dH2O 1ml 5 Lysis Buffer 625 ul x 4 6 Proteinase K 20 mg ml 50 ul 7 dH20 for Dilution 150 ul 8 Giardia Positive Control 2 1 x 105 copies ul 50 ul 9 EASY Dilution for Real Time PCR 1mx 2 1 2
23. Error http www takara bio coJp 13 CY231 Bi 1 OUT 2 DNA OUT FAM ROX PCR ROX FAM gt Giardia Primer Probe Mix
24. 18 1 Ml 2 G PCR 4 RNase Free dH2O Giardia Positive Control J PCR J 5 VI 1 n 2 1 1 o 0 2 ml 1 5 X PrimeScript RT Master Mix 2 ul 1 Xx 1 5 u RNase Free dH20 up to 10 ul 10 ul RNase Free dH20
25. 1 1 0 2 ml 1 5 X PrimeScript RT Master Mix 2 L 1 Xx 2 ul RNase Free dH2O 6 ul Total 10 ul 2 3 Thermal Cycler 37 15 85 5 4 C 20 http wwwr takara bio coJp 18 CY231 VII 2 PCR 1 18S RNA DNA
26. 02 ml PCR 1 http wwwr takara bio coJp 9 CY231 VII 3 PCR 3 PCR Thermal Cycler Dice Real Time System Applied Biosystems 7500 Fast Real Time PCR System Life Technologies Thermal Cycler Dice Real Time System 1 gt OK RSAT DESIA Hesg pika ki
27. DNA FAM 18SrRNA ROX DNA DNA IC PCR DNA DNA PCR DNA PCR DNA DNA
28. Omit Set Omit gt Reset 6 1 2 3 4 CP Crossing Point 5 6
29. http wwwr takara bio coJp 24 5745 E CY231 gt 23 23 45 1 3GTIn x 39 363 R 0 9989 40 35 3o 25 x 20 5 L 1 424ln x 29 262 R 0 9079 ctf 10 4 5 0 r i 1 00E 05 1 00E 03 1 00E 01 1 00E 01 1 00E 03 1 00E 05 or CopiestqPCR 9 1 18S rRNA 1 600 PC Giardia Positive Control
30. Ther mal Cycler Dice Real Time System Software lt PM M Plus Minus Assay gt Thermal Cycler Dice Real Time System 2 PCR 2 Step PCR Hold Cycle 1 96 30 2 step PCR Cycle 50 96 10 60 20 HAER 96 0 60 0 O f gt http www takara bio coJp 10 CY231 3 4 ROX Omit http www takara bio cojp 11 CY231 5
31. PCR 1 4 PCR 1 4 5 X PrimeScript RT Master Mix Buffer dNTP Mixture 2 X Cycleave Reaction Mixture Giardia PCR Buffer dNTP Mixture Giardia Primer Probe Mix DNA DNA 18S rRNA
32. Giardia Positive Control G ND FEAM ROX http wwwr takara bio coJp 14 CY231 Applied Biosystems 7500 Fast Real Time PCR System Life Technologies Ver 2 0 1 Experiment Properties Quantitation Standard Curve TaqMan Reagents Other 2 Plate Setup Define Target Target name 2 Giardia Reporter FAM Quencher None IC Reporter ROX Quencher None 3 Define Sample NCG Positive Control 4 Plate Layout Passive Reference none Deiro Taqet ang Sarnos Me and Smplos PUNUE D IA a tirta ahta eat le
33. 18S rRNA DNA OD26o 1 x 10 copies 1 EASY Dilution for Real Time PCR Giardia Positive Control 10 1 Ml 2 3 PCR 4 J Giardia Positive Control J RNase Free dH2O PCR 4 5 VIII 1 n 2
34. ROX IO FAM ROX target http www takara bio cojp 16 CY231 VII 4 1 ROX 2 3 2 Giardia Positive Control ROX FAM 4 3 3 RNase Free dH2O ROX 3
35. http wwwr takara bio coJp 19 CY231 VIII 3 PCR 3 PCR Thermal Cycler Dice Real Time System Thermal Cycler Dice Real Time System 1 OK 5347 Enea vj ezk MEROE az aew Thermal Cycler Dice Real Time System Software AQ M Absolute Quantification gt 2 PCR 2 Step PCR 1 2 Hold Cycle 1 96 C 30 2 step PCR Cycle 50 96 C 10 60 C 20
36. 0302 2 4 3 10 20 L a PTFE 10 ml Dynabeads GC combo VT j PCR RS
37. 1 1 EWL 7 n ra EE RRR Prey Cyw e E 24 4 amp 10 12 ME 20 22 34 26 13 12 W 2 40 42 44 4i6 12 M4 HMR mesane Troe sc fe G ss 2 6 ef E oe j HE RT 2 9 8 19 12 t4 9 8 29 22 34 2 20 39 32 34 3 30 4 42 4 4648 99 7472 FAM ROX IO j Elk Auto 2 NC PC Gg 7a Positive Control NC N gt FAM ROX FAM ROX IC o 0 4 6 816112 14 10 1620 23 24 24 20 10 13 14 WW 40 EE TE 2 4 6 1 t2 14 t4 18 20 22 2 25
38. DNA RNA cDNA DNA RNA RNA PCR RT PCR PCR PCR RNA DNA RNase H
39. 4 28 10 i3 3M 26 38 4D 42 44 4 AB S0 4250 Aran lal E DEBR mey Sw 6 88 http www takara bio coJp 12 CY231 PC P gt PC FAM FAM ROX IC e o m n i TT i t 20 i 4 AG MN HG 16 202224 24 48 WD MMI 40 4 gt 44 4E 44 54 a 3 4 6 81 t2 14 t T8 2 22 2i 24 28 3i0 aj 3M 24 38 40 42 44 tt ror tinan 3 gt 6 gt an e o HATITar J C A N gt P gt OK OUT gt Posi Nega ND PCR
40. e r CY231 IdfaR4 Cycleave RT PCR Giardia 18S rRNA Detection Kit v201402Da Giardia lamblia G intestinalis G duodenalis 5 18S rRNA
41. f 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 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 M NANOGEM 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 Techno
42. logy 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 coJp 27 CY231 TafdR4 Tel 077 543 6116 Fax 077 543 1977 http www takara bio coJp JND TIRS v201402Da
43. pdt se gr et inewh Y t Semi bart wmitrm TH Ez m 5 Instrument Pi hag FPPernand 0 227 p loe handan Ora norner Hold Cycle 1 96 30 2 step PCR Cycle 50 96 10 60 C 25 25 KK te a http www takara bio co jp 15 CY231 6 Start 7 Analysis Amplifcation Plot Threshold Baseline Manual FAM IC ROX we km mm we k m 8 View Well Table View Well Table gt View Piate Layout Selact Wells Wh Salectitam lt Sampe TWIN_ Task Standard Curva Muancomponen Piot Raw Data Plot A Summary Mapio Phris View gt StepOnePlus Real Time PCR System Life Technologies ROX target
44. r Real Time NJ500 Plate Sealing Pads 9090 0 2 ml Hi 8 Tube NJ300 0 2 ml Hi 8 Flat Cap NJ302 http wwwr takara bio coJp 26 CY231 Epoch Biosciences nc
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