Cycleave®PCR EHEC (O165)
Contents
1. 5 PCR PCR Auto PCR Manual http wwwr takara bio coJp 4 CY241 V 1 DNeasy Blood amp Tissue Kit DNA 2 PCR 3 PCR 4 4 1 22. PCR 1 http www takara bio coJp 6 CY241 4 3 PCR 3 PCR Thermal Cycler Dice Real Time System Quick Manual gt PCR Hold Cycle 1 95 10 3 step PCR Cycle 45 95 5 55C 10 726 20 FAM lt
3. VII dH2O 0165 Positive Control V 0020 0165 Positive Control http www takara bio coJp 12 CY241 VII Cycleave PCR EHEC O157 O26 Typing Kit CY237 Cycleave PCR EHEC O111 O121 Typing Kit CY238 Cycleave PCR EHEC 0103 0145 Typing Kit CY239 Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice Real Time System Lite TP700 TP760 EASY Dilution for Real Time PCR 9160 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 gt Cycleave PCR 0 157 VT gene Screening Kit Ver 2 0
4. Rev ne mast kr nyyas iP lp Te Fs At Wnt tiqk Tet Deut 19 tf Eee sm tl Tr oC NE on Te ie 386660 Ust nan hetgs he IN GOCte De Cl CT NH TMR mh Ref TO MITO SRpeneaww Soas las hfemneO Swine Cons mmm 1 Toc Time 12900 0 Sadcwie 28 EwdC mis 272 Am apcn Pt Threshold i oo oo Baseline et oot 0201 00001 060OCO1 0000007 2 Cycle Options Target 0 Tmwsshold auto 2000 show Threshoid 22898158 Start Weil WTarget 2 9386108 5nd WeW Target StepOnePlus Real Time PCR System Life Technologies http wwwr takara bio coJp 10 CY241 VI lt A gt
5. DNA DNeasy Blood amp Tissue Kit QIAGEN DNeasy Blood amp Tissue Kit NucleoSpin Tissue 740952 10 50 250 V 2 2 lt A gt O DNA B PCR gt PCR 1 100 Positive Control DNA B gt A gt 0O165 VT V 1
6. 0 DNA dH2O 0165 Positive Control B PCR O165 B PCR lt B PCR gt PCR 1 100 Positive Control DNA lt A gt O165 dH2O PCR PCR 0165 FAM dH20 PCR http wwwr takara bio coJp 11 CY
7. 1 3 4 5 2 X Cycleave Reaction Mixture PCR Buffer dNTP Mixture 0165 Primer Probe Mix 0165 0 0165 FAM dH20 0165 Positive Control O165 EASY Dilution for Real Time PCR Positive Control 206 http www takara bio coJp 3 CY241 PCR Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice Real Time System Lite TP700 TP760 Applied Biosystems 7500 Fast Real Time PCR System Life Technologies DNeasy Blood amp Tissue Kit QIAGEN
8. Threshold Baseline Manual 1 Amplification Plot Options Target 0165 Target Threshold Auto Auto Baseline Show Threshold Baseline Rn 01 ODL TET sumple 2 1 0 wlTmeshoid 00001 ue 8339106 ow Tmmesnoa Boseine wellg Target 8 End W Target 2 Analysis Setting Analysis Settings analysis Settinds 6 http www takara bio coJp 9 CY241 3 Analysis Setting Select a Target O165 Target Ct Settings for 7arget Name Threshold Baseline Apply Analysis Settings Threshold Amplification Plot Log Baseline Amplification Plot
9. PCR B gt 1 Positive Control lt B gt 3 0165 Positive Control EASY Dilution for Real Time PCR 1 100 0165 Positive Control 1 0165 Positive Control 5 ul EASY Dilution 45 AI 1 10 2 1 1 10 5 EASY Dilution 45 ul 1 100 http wwwr takara bio coJp 5 CY241 2 1 lt A gt o 20 ul 1 dH2O 5 2 O165 1 2 x Cycleave Reaction Mixture 125 ul 10 0165 P
10. RNase H RNase H RNA RNA 1 http www takara bio coJp 2 CY241 25 Il 50 1 2 X Cycleave Reaction Mixture 2 O0165 Primer Probe Mix 3 dH20 4 0165 Positive Control 5 EASY Dilution for Real Time PCR 2 conC 5 Xx ConC 625 250 HI 1 ml 100 ul 20 1 ml
11. CY217A B EHEC VT gene PCR Screening Set RR120A IX Cycleave Thermal Cycler Dice TaqMan Roche Diag nostics GmbH http wwwr takara bio coJp 13 CY241 http wwwr takara bio coJp 14
12. 200 HU 20 MI 10 ul IV 1 PCR 2 3 4 3 1 2 3
13. CY241 http wwwr takara bio coJp 15 CY241 TafdR4 Tel 077 543 6116 Fax 077 543 1977 http www takara bio coJp v201408Da
14. 241 A O165 PCR B PCR PCR DNA B 0165 PCR Positive Control
15. gt lt none gt NC Negative Control PC Positive Control UNKN Unknown http wwwr takara bio coJp 7 CY241 Applied Biosystems 7500 Fast Real Time PCR System Applied Biosystems 7500 Real Time PCR System Life Technologies Advanced Setup Experiment Properties Quantifcation Standard Curve TaqMan Re agents PCR Hold Cycle 1 95 10 3 step PCR Cycle 45 95 5 55C 10 726 25 Passive Reference none Define Targets Target Name 0165 Reporter FAM Quencher none Define Samples NTC No Template Control Unknown Manual http wwwr takara bio coJp 8 CY241 Analysis Amplifcation Plot
16. ma Y 4 IdfaR4 Cycleave PCR EHEC 0165 Typing Kit vV201408Da 0157 H7 026 EHEC EHEC EHEC M O O O165 O165 O FAM PCR RNA DNA
17. rimerProbe Mix 5 ul 1 X or 0165 Positive Control 5 ul or dH20 6 2 5 ul Total 25 ul lt B PCR gt o 20 ul 1 dH20 5 1 1 2 x Cycleave Reaction Mixture 12 5 ul 1 X 0165 PrimerProbe Mix 5 ul 1 O165 Positive Control 1 100 0 5 MI or dHzO 5 ul O dHO 20 Total 25 ul A gt DNA 3 3 3
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