Cycleave®PCR Legionella Detection Kit
Contents
1. IV 1 5s rRNA TOREA ne KETC 12064 K5951 700510 uma 700560 gt ueo 700562 1 NgO417 NIIB0406 L drozanskii NIIB0078 L dumoffi 14 NIIB0049 L erythra NIIBO146 L fairfieldensis NIIBO408 L fallonii NIIB0417 L donaldsonii 1 2 1 1 3 12 13 15 1 1 1 6 7 8 9 http www takara bio co jp 4 CY209 on Seis ooro AMET a rar noar coana E m0 fom 22 es zgeegs E em roo 3 Weoeeolz7egeee mg E 4 eoee i hackelae 2 lt mAm aso 25 Weooss seess seceww4 ang gt 26 eooe Ljamestownienss E Azeerg as gt ar noa jordans SSC E sese ea 9 nmorao ensinos lt igwm aos 29 ez enews 1 E swl 30 en25s ondinensis 2 E Mulhouse 26 maa 32. PCR HAEDH 1 47 mm 47 mm 0 22 um 2 500 ml 3 50 ml 4 5 ml 1 5 2 ml 2 ml 6 13 000 15 000 rom 4 5 AIDA RE ER 950 ul x 2 ZIP 100 ul http www takara bio co jp 7 CY209 DNA MonoFas DNA 1 NN NW BW N SW Ws wes es gt O O
3. RNaseH RNA 1 o RNA RNase H ote ee DEE 1 http www takara bio co jp 2 CY209 lL 25 ul X 50 1 TaKaRa Ex Tag HS 5 units u 12 5 ul 50 2 Tli RNase H II 200 units u l 25 ul 50 3 5 X Reaction Mixture 2 5 X conc 250 ul 50 4 Primer Mixture 150 ul 50 5 Probe Mixture 3 150 ul 50 FAM TET ROX 6 Mip Positive Control 10 ul 10 7 5S Positive Control 10 ul 10 8 dH20 1 0 ml 1 Tli RNase H Il t Thermococcus litoralis RNase H 2 dNTP mixture Internal control 3 GCM Probe 8
4. ER pneumophila 1 L pneumophila 10 0 5 CFU 5 Xx 104 CFU ul 6 2 ul 2 IVIL PCR dH20 Negative Control 3 PCR FAM TxR TET mip FAM sample IE ANTE Texas Red Temperature Standard chi Standard ch3 Views Results Tale Analysis Settings Protocols Feu TET Texas Red Temperature Standard chi Slandard ch3 TET Temperature Standard chi Standard ch3 http www takara bio co jp 19 CY209 4 Ct
5. Scale graph Axes Graph Scale 2 y Axis FI Max Y AJL Apply 3 Exit Axes Graph Scale http www takara bio co jp 21 CY209 XI 1 Fields B S Benson R F Besser R E Legionella and Legionnaires disease 25 years of investigation Clin Microbiol Rev 15 506 526 2002 2 Benin A L Benson R F Besser R E Trends in legionnaires disease 1980 1988 de clining mortality and new patterns of diagnosis Clin Infect Dis 35 1039 1046 2002 3 1999 4 19 FE
6. A Bo pal C Tel 077 543 6116 Fax 077 543 1977 NANOGEN http www takara bio co jp http www takara bio co jp 22 CY209 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 a
7. Epoch Biosciences Inc Epoch Biosciences Inc US20020155484 WO0142505 WO02099141 US10 113 445 US09 876 830 Epoch Biosciences Inc Epoch Biosciences Inc
8. mip FAM Log Concentration Standard chi Standard cha 4 a 19 21 23 25 ar 20 31 33 35 a 30 Threshold Cycle y 0303x 12 127 requared 0 996 Results Table Analysis Settings Protocols E FAM TET Texas Red Temperature standardehi standard cha Log Concentration 48 20 22 24 26 28 3 32 34 36 33 40 Threshold Cycle y 0 304x 11 843 requared L pneumophila 10 CFU http www takara bio co jp 20 CY209 X Allogrescence a a Press FA keyto reset zoon Ctrl A to show all sites Chick right mouse buttondo scale grap FAM TET TxR Std Res POS NEG Result Table Sample Type STD Sampl
9. http www takara bio co jp 8 CY209 Vil 1 Smart Cycle ll 3 Smart Cycler System Smart Cycler Il System Smart Cycler gt Smart Cycler System ChI Ch 2 Ch 4 Manual Thresh Fluor Units 100 16 17 1 Smart Cycler Il 2 Protocol Define Protocols New Protocol Protocol Protocol 2 e Smart Cycler User Logs Setup Tools Help e New Probocod Please enter anew name forthe Protocol Cena Protocol Stage 7 Repeat 40 times DegSec ITI Secs Optics NA 55 110 O New Plotocol NA 720 6 On _ Advance to Hert Stage http www takara bio co jp 9 CY209 3 Graph Graph
10. XII Smart Cycler Il System SC200N Cycleave PCR Legionella 5S rRNA Detection Kit Ver 2 0 CY210 Cycleave PCR O 157 VT1 VT2 Typing Kit CY222 Cycleave PCR 67 CY218 Cycleave PCR Salmonella Detection Kit Ver 2 0 CY205 XIII
11. 100 cfu tube DNA 10 000 cfu tube DNA E morelense Legionelagenomospecies T NIIB0197 L worsleiensis 61 62 63 64 65 67 70 71 72 73 74 75 19 2 mip L pneumophila sero group 1 2 3 4 5 6 7 10 3 mip L pneumophila worsleiensis L fairfieldensis 4 Serratia marcescens Alcaligenes faecalis Klebsiella pneumoniae subsp pneumoniae Staphylococcus epidermidis Achromobacter xylosoxidans subsp denitrificans Streptcoccus pyogene
12. RIDE http www takara bio co jp 13 CY209 Smart Cycler Start Run IR A pm twee co anja PEA lee rr Por hie FY Linea FR Lie Pa Lae PA Pii Lisa PE EmN 1 PLA Lar Liar PON La hi ap ol hay tl Lora Pa Te Gg ee TARA a A AMA A EE Eph La mira cle 4 aa Pra HeH wi ro 4 a FTP Haah at a F i CE Pom lore ieii fa Ta a or 4 an Por Dero tal 20 ud Fa L hi Start Run 5 1 View Results View Results View Results 2 Select Graphs Select Graphs FAM mip TxR 5S rRNA TET Temperature Select Graphs Select Graphs mi Heys BYHR Green Meli X FAM TET Texas Re
13. eg o e7g bo Oe Ce x2 POS 3 PCR 4 5 http www takara bio co jp 17 CY209 Mip Positive Control 5S Positive Control Ch 1 mip FAM http www takara bio co jp 18 CY209 IX CFU
14. DNA 3 1 100 Hot Start PCR 3 Takaka Ex Tag HS 2 age RNA DNA RNase H RNA
15. Std Res http www takara bio co jp 16 CY209 2 Mip 55 Positive Control BA TET Std Res FAM mip mip Std Res mip mip 4 3 5S rRNA 5S rRNA nes 5S rRNA 4 3 3 TET Std Res HA E POS mip mip pa 5 es mip NEG mip e E eo 3 POS 5S rRNA 5SrRNA TxR ERA 5 RELY 5 Std Res 55 rRNA k1 POS NEG mip 5S rRNA pneumophila
16. CY209 IdRaRa Cycleave PCR Legionella Detection Kit v201308 Legionella pneumophila IFA 70 E Legionella pneumophila dd CycleavePCR Legionella Detection Kit PCR pneumophila
17. FAM Std Res POS 100 NEG 5S rRNA TxR Std Res TET Std Res Views 4 Site gaa SamplelD Sample Notes Statug FAm Ct Results Table ID Type Analysis Settings HAB Legionel NTC UNKN OK doo Protocols JAD Legionel 1 CFU UNEM OK LU FAM HATO Legionel 10 CFU UNEN OK 6 90 TET HATI Legionel 10 2 GFU UNEN OK 354 Texas Rad 412 Leglnngl 10 3 CFU UNE OK Temperature JAI Legionel 1074 CFU UNEN HATA Legionel 1075 CFU UNEN OK i mip 5S rRNA x FAM td Res TXR Std Res POS NEG 21 X 1 Vill mip FAM 100 Results Tables FAM Std Res POS 100 NEG 5S rRNA TxR ctd
18. 5010 21555 gt 99 70 http www takara bio co jp 3 CY209 ll 20 C Il 1 2 ews 3 HE 121 20 2 5
19. 2 3 VI 2 90 PCR
20. Dye Set a name Sub Min Cycle Max Cycle FA Assay OH 40 Primary Cura Manual Protocols 42 TET Assay JON 40 Primary Cure Manual Protocol Lnthumbem PR Assay ON 5 HH PrimaryCure Manual a CyS Assay OM j5 40 Primary Cure Manual Select Protorode ood es Site Protocol OK Select Protocek and Gites Le PGR Loponen PER Leroa PER Lepom PCR E Lepore PCR Logia PCR Logica PEA http www takara bio co jp 12 CY209 2 PCR 1 Mip Positive Control 5S Positive Control Mip Positive Control 5S Positive Control 1 Positive Control 1 o 1 5 X Reaction Mixture 5 ul Primer Mixture 3 ul Probe Mixture 3 u
21. FOIE Smart Cycler Il System Smart Cycler System pS FAM ye ROX TET Eclipse ome ne Eclipse Epoch Biosciences lt gt PCR Smart Cycler System Smart Cycler Il System SC200N Smart Cycler SC910C Smart Cycler SC900 1 000 ul 200 ul 20uk 10 lt DNA gt 47 mm 47 mm 022 um Millipore Code HTO4700 Isopore 50 ml 2 0 ml MonoFas DNA
22. 2 3 1 mip channel 1 FAM Define Graphs Save Graph FAM Stop Run Automatically added to new Runs Graph Type Optics y Ehannel sk 4 Ch 1 Ch2 _ cha 7 cha r Show el Primary Curve 2nd Derivative I Threshold Horizontal _ Threshold Crossings Vertical Axes w Auorescence vs Cycle gt Log Fluorescence ve Cycle 3 2 5S rRNA channel 3 Texas Red ROX Texas Red channel Ch 3 Texas Red http www takara bio co jp 10 CY209 3 3 channel 2 TET TET Stop Run _ Automatically added to new Runs Graph Type Optics gt Channel s i
23. Weoesz z ogeectee 1 lt tong each 33462 gt 732 Weoegs Longbeachae 2 lt Tucker os eooes imacete E meg aso 3a eoogslzaceegg lt mmow ame gt 3s nwon emana g sess EZ P36 nos inann g im E 37 NWeoosslzoeegess g ori EE 39 eoog e parsers E przeco as gt 2 c mm aise 3 emigra as 42 NiB0004 L pneumophila subsp fraser 4 Cc tosAngeles1 33156 3 NiB0005 pneumophila subsp frase 5 E Oalasie 33216 E ETE goat 5 ww EA E neo e s lt qewz sees c cweei E e neno e E waero asa PP o lt emi Jem iB DS 4 min m L pneumophila subs NIIB0060 Sheumophila j 570 CO H 43290 L pneumophila subs L pneumophila subs NIIB0451 L pneumophila untypable E H13 192 NIIB0453 L pneumophila untypable E H13 206 NIIB0462 L pneumophila untypable E H13 239 E 1 http www takara bio co jp 5 CY209 i Bi RARE oe e a Nioo Laban it e Neono sespe US i n ae a Legionella genomospecies 1 A NIIB0306 Legionella sp E LLAP 14 700313 10 cfu tube DNA 10 cfu tube DNA
24. Ch 1 mi Ch2 ch3 chs ivi Primary Curve 1 2nd Derivative Threshold Horizontal J Threshold Crossings Vertical Fluorescence vs Cycle Log Fluorescence vs Cycle E 4 Create Run Run Name Dye Set FTTC25 Smart Cycler User Logs Check Status Stop Ron GI Site ge Protocol Sample ID Sample Type Note Run Name di lLegionetta POR TEST i Notes AddRemove Sites TREES a E AI PE gS ear rt FOR CoE OR TOA Ch Dye Usage Bkand Bkgnd Bkgnd Curve Analysis Thresh Mame Sub Min Cycle Max Cycle cM FAM Assay ON 5 40 Primary Curve Manual Hz Cy3 Assay ON 5 40 Primary Curve Manual Ja TR Assay ON 5 40 Primary Cure Manual ofe CV5 Assay ON 5 lso Primary Curve Manual Dye Set http www takara bio co jp 1 1 CY209 5 Add Remove Sites Select Protocols and Sites Site Protocol OK Smart Cycler User Logs Setup Tools ele Sig Aun O a ak Sample Typ mr om Legionella POR TEST A E Select Protocols and Sites ane LE a ea Dye Usage Bignd Bkgmd kond CURE AEAN Tesh
25. CR Licensed under U S Patent No 5 338 671 and 5 587 287 and corresponding patents in other countries 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 23 CY209 Takaka Tel 077 543 6116 Fax 077 543 1977 http www takara bio co jp STII 7A ARS 4 v201308
26. O 100 ul Buffer Mix 90 40 15 Proteinase K 10 60 1 75 5 20 000 X g 15 000 rpm 3 Buffer AL 200 ul 200 pl MonoFas Column for Legionella genome 2 300 xg 5 000 rem 1 Buffer AW1 1 300 9 000 X g 10 000 rpm 1 Buffer AW2 300 ul 0 L 15 000 x g 13 000 rpm 2 Buffer AE 20 ul 1 15 000 X g 13 000 rpm 1 DNA 5 ul PCR 1 MonoFas DNA
27. Res TET Std Res FAM Std Res TxR Std Res POS gt 21 X 2 1 TET Std Res mip mip l L pneumophila L pneumophila t es mip lt ieee pneumophila 5S rRNA 1 5S rRNA 1 Legionella Legionella 5S rRNA 2 3 Legionella
28. d Temperature http www takara bio co jp 14 CY209 3 Views Results Table Sample ID Sample ID _ Views Site Protocol SamplelD Sample Results Table LD Tyee Analysis Settings phl Leglonella_PCRANT AT iM Protocols AZ Legionella PCR INEN Faw IA3 Legionella_PCR NEN TET 44 Legionella PER 10 MINK Texas Red IA5 Legionella PCR 10 3 CFU NIN Temperature JAG Legionella PCR 10 4CFU UNEN AA Legionella PER 10 5 CFU UME 4 Analysis Settings Ch 4 Usage Unused Ch Results Table Ch 1 2 3 Assay Ch Dye Usage Bkond Name Sub Min Cycle Max Cycle 1 FAM Assay ON 5 140 Primary Curve Manual 10 12 TET Assay ON 5 40 Primary Curve Manual At TR Assay ON 5 40 PrimaryCurve Manual 10 44 Cy5 L ION 55 40 Primary Curve Manual 30 Texas Red dl Temperature S unused ER 5 Analy
29. e Type UNKN POS Press R Hey to reset zoom Ciri Ato show all ates Click right mouse bution to scale graph LE Cycles a HI ETICE Press R Hey to reset room Ciri A to show all sites Click right mouse bution to scale graph Y Y Smart Cycler System 1 Y Axes Graph Scale 2 y Axis Fl Max Y 3 Exit Axes Graph Scale Smart Cycler Il System 1 Y
30. l TaKaRa Ex Taq HS 5 U ul 0 25 ul Tli RNase H II 200 U ul 0 5 ul Positive Control 5 ul TUS 3 dH20 8 25 ul 25 ul E Positive Control 1 ul G2ul 3 5 ul 20 ul El 3 3 3 1 2 Positive Control 4 PCR 3
31. mip macrophage infectivity potentiator protein pneumophila 5S rRNA PCR Smart Cycler System Smart Cycler Il System mip FAM 5S rRNA ROX TET Smart Cycler System Smart Cycler Il System 3 1 mip 5S rRNA 30 x Smart Cycler Cepheid 1 PCR PCR Ald CK DNA DNA
32. nd 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 Il 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 USO9 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 MN NANOGEN L15 Hot Start P
33. s Pseudomonas fluorescens Pseudomonas aeruginosa Pseudomonas stutzeri Acinetobacter baumannii Acinetobacter Iwoffii Myroides odoratus Escherichia coli 0157 H7 Salmonella Enteritidis Salmonella Agona Salmonella Typhimurium Salmonella Dublin Salmonella Infantis Salmonella Arizonae Citrobacter freundii Morganella morganii Staphylococcus aureus enterotoxin A B C D E F G H human genome http www takara bio co jp 6 CY209 V je Smart Cycler System 2 PCR 3 1
34. sis Settings Ch 1 2 3 Manual Thresh Fluor Units 100 Cut Off Update Analysis Bkond Bkond Cure Analysis Thresh Setting Manual Thresh n he Min Cycle Max Cycle Fluor Units 1 FAM sy ON 5 40 Primary Cure Manual 100 0 ITET Assay ON 5 40 Primary Cume Manual 00 0 Manual Thresh T Assay ON 5 40 Primary Curve Manual 100 0 ICy5 JUnus ON 8 AD FrimayCume Manual 30 0 Fluor U nits 100 Texas Red Temperature 6 Views FAM mip 7 Views Texas Red 5S rRNA 8 Views TET http www takara bio co jp 15 CY209 9 Results Table FAM Std Res Standard Results TxR Std Res TET Std Res mip ee 100
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