SYBR® Premix Ex Taq ™ II （Tli RNaseH Plus）
Contents
1. 2 3 SYBR Green 4 http wwwr takara bio coJp 4 RR820A VI lt Thermal Cycler Dice Real Time System gt X Thermal Cycler Dice Real Time System Perfect Real Time PRTSS 1 PCR lt 1 gt SYBR Premix Ex Taq II Tli RNaseH Plus 2 X 12 5 ul 1x PCR Forward Pri2. TaKaRa Ex Tag H5 7ag PCR PCR PCR 95C 5 15 PCR 95 30 3 http www takara bio co jp 5 RR820A PRTSS 1 PCR lt 1 gt SYBR Premix Ex Taq Il Tli RNaseH Plus 2 x 12 5 ul 1X PCR Forward Primer 10 uM 1 ul 0 4 uM 1 PCR Reverse Primer 10 uM 1 ul 0 4 uM 1 template lt
3. SYBR Premix Ex Taq 1 Tli RNaseH Plus RR820S A B SYBR Premix DimerEraser Perfect Real Time RR091A B SYBR Premix Ex Taq Tli RNaseH Plus RR420S A B at SYBR Premix Ex Taq SYBR Premix Ex Taq Il SYBR Premix DimerEraser http wwwr takara bio coJp 13 RR820A VII Appendix 1 PCR
4. RT PCR amp Perfect Real Time http www takara bio co jp realtime RefSeq RT PCR SYBR Green RT PCR RT PCR PCR 6 12 80 150 bp 300 bp 17 25 mer 40 60 45 55 Tm Forward primer Reverse primer Tm Tm OLIGO 1 63 68 C Primer3 2 60 65 C
5. PCR VII 1 RT PCR amp Perfect Real Time http wwwr takara bio coJp realtime http www takara bio coJp 3 RR820A Ill 4 6 80 20 4 6 IV 1 PCR 2 SYBR Green
6. O No Template Control O Ct PCR 100 0 2 uM 1 0 uM A http wwwr takara bio coJp 12 RR820A PCR O
7. 1 PCR lt 1 gt GAs SYBR Premix Ex Taq Tli RNaseH Plus 2X 10 ul 25 ul 1x PCR Forward Primer 10 uM 0 8 ul 2 ul 04 uM PCR Reverse Primer 10 uM 0 8 ul 2 ul 04 uM 1 ROX Reference Dye 50 X or Dye ll 50 x 2 0 4 ul 1 ul 1x template 3 2 ul 4 ul dH2O 6 ul 16 ul Total 20 ul 4 50 ul 4 1 primer 0 4 UM 0 2 1 0 UM 2 ROX Reference Dye II 50X ROX Reference Dye 50X Applied Biosystems 7500 7500 Fast Real Time PCR System ROX Reference Dye Il 50x StepOnePlus 7300 Real Time PCR System ROX Reference Dye 50 x 3 template 20 ul DNA template 100 ng RT PCR cDNA RT template
8. SYBR Molecular Probe Inc http wwwr takara bio coJp 17 RR820A NOTICE TO PURCHASER LIMITED LICENSE L11 SYBR Green This product is covered by the claims of U S Patent No 5 436 134 and 5 658 751 and their foreign counterpart pat ent claims Takara PCR products containing SYBR Green are sold under license from Molecular Probes Inc only for the usage in Real time PCR for internal research purpose These products are not to be used for the purpose such as provid ing medical diagnostic or any other testing analysis or screening services or providing clinical information or clinical analysis in return for compensations L15 Hot Start PCR Licensed under U S Patent No 5 338 671 and 5 587 287 and corresponding patents in other countries L46 SYBR Melting Curve Analysis The purchase of this product includes a limited non transferable license for all fields other than human or veteri nary in vitro diagnostics under specific claims of U S Patent Nos 6 174 670 6 569 627 and 5 871 908 owned by the University of Utah Res
9. PCR Sample volume 25 ul Step1 95 C 30 Step 2 PCR GOTO 39 40 95 5 60 30 Step 3 Melt Curve TaKaRa Ex Taq H5 7ag PCR PCR PCR 95 C 5 15 PCR 95 30 3 CFX96 PCR http www takara bio coJp 7 RR820A lt Applied Biosystems 7300 7500 7500 Fast Real Time PCR System StepOnePlus Real Time PCR System gt
10. PCR 95C 5 95C 5 60 30 64C 30 O 3 step PCR PCR 3 step PCR 3 step PCR as loc 5 S195 5 95 5 95 C 5 1 60 C 30 60C 1 55 C 30 55 C 30 72 C 30 72C 1 95 30 DNA 95 1 2 2 SYBR Green Assay PCR 3
11. RefSeg RT PCR SYBR9 Green RT PCR X 1 2005 3 4 PCR 120 126 2 2008 PCR 3 1 RT PCR 39 43 Xl
12. 100 PCR 7aKaRa Ex Taq HS http wwwr takara bio coJp 2 RR820A 2 DNA SYBR Green DNA DNA 1 F LL LL tt pt 2 OF F eM F F F cys ee DD DD S E S S A A A A 3 oe gt M DO OO OO ALP LLLLLLIL
13. Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice Real Time System Single TP850 TP870 Thermal Cycler Dice Real Time System Lite TP700 TP760 Applied Biosystems 7300 7500 7500 Fast Real Time PCR System StepOnePlus Real Time PCR System Life Technologies 4t LightCycler LightCycler 480 System Roche Diagnostics CFX96 Real Time PCR Detection System Bio Rad G Smart Cycler System Smart Cycler Il System Cepheid SYBR Premix Ex Taq Tli RNaseH Plus RR420S A B Molecular Probes Inc SYBR Greenl SYBR Molecular Probes Inc 7aKaRa Ex Taq Hs PCR PCR SYBR Greenl 1 PCR PCR DNA DNA DNA 3 1
14. 2Xconc PCR 3 PCR PCR ABER Takaka Ex Taq Hs PCR 4 2 RNase H Tli RNase H cDNA mRNA PCR V 1 SYBR Premix Ex Taq II Tli RNaseH Plus 2Xconc 80
15. Dissociation 4 PCR 12 TaKaRa Ex Taq HS 7a9 PCR PCR PCR 95C 5 15 PCR 95 30 3 http www takara bio co jp 6 RR820A CFX96 PCR gt CFX96 PCR Bio Rad 1 PCR
16. total RNA1 pg 100 ng cDNA Negative Control dH20 http www takara bio coJp 15 RR820A VII O Thermal Cycler Dice Real Time System human Testis CDNA PCR 185 bp SYBR Green TaKaRa Ex Taq HS O DNA 74 30 10 nmol 1U O 25 mM TAPS pH9 3 25 50 mM KCI 2mM MgCl2 0 1 mM DTT 200 uM dATP dGTP dCTP 100 u M BH dTTP 0 25 mg ml DNA O 1 10 U 0 6 ug O A Hind Ill DME 74 C 1 DNA 2 10 U 0 6 Ug supercoiled pBR322 DNA 74 C 1 DNA 3 10 U 0 6
17. GC AT 3 T C polypyrimidine A G polypurine 3 3 GC AT 3 G C 3 T 3 base 3 2 base BLAST 3 1 OLIGO Primer Analysis Software Molecular Biology Insights 2 Primer3 http www genome wi mit edu ftp distribution software 3 http www ncbi nlm nih gov BLAST X Tik Perfect Real Time TEL 03 3271 8553 TEL 077 565 6969 http wwwr takara bio coJp 14 RR820A 2
18. PCR 95 C 5 15 PCR 95 30 3 http www takara bio co jp 11 RR820A PCR PCR SYBR Premix PCR RR420S A B RR091A B
19. RT PCR RT PCR PrimeScript RT reagent Kit Perfect Real Time RR037A B PrimeScript RT Master Mix Perfect Real Time RRO36A B PrimeScript RT reagent Kit with gDNA Eraser Perfect Real Time RR047A B 1 PCR Thermal Cycler Dice Real Time System o 22 5 24 ul 1 gt SYBR Premix Ex Taq ll 2X 12 5 ul 1X PCR Forward Primer 10 uM 1 yl 0 4 uM PCR Reverse Primer 10 u M 1 yl 0 4 uM dH2O X Ul Total 22 5 24 ul 2 1 2 5 PCR 2 5 PPPS SF SCPMMUAVKSHNTMUBMSBIMHBAUMNBAAKHRUKASEHIUAR HB ore KO al ti ret 5 Se WSWO ieee ee ee 2 R2 1 000 5 Eff 101 9 a I 13 1 10 xe EXO Oe RT PCR Human TBP mRNA
20. lt 1 gt SYBR Premix Ex Taq Il Tli RNaseH Plus 2X 12 5 ul 1x PCR Forward Primer 10 uM 1 ul 0 4 uM PCR Reverse Primer 10 uM 1 ul 0 4 uM template lt 100 ng 2 2 ul dH2O 8 5 ul Total 25 ul 1 primer 04 UM 0 2 1 0 uM 2 template DNA template 100 ng RT PCR cDNA RT template PCR 10 2 PCR PCR PCR Tm PCR 3 PCR PCR 12
21. PCR 10 4 2 PCR PCR PCR Tm PCR 3 PCR PCR 12 lt Applied Biosystems 7300 7500 Real Time PCR System StepOnePlus gt PCR Stage1 Stage2 Stage 3 Stage 1 Reps 1 Reps 40 Reps 1 Reps 1 0 95 C 30 Stage 2 PCR Reps 40 95 C 5 60 30 34 Stage 3 Melt Curve StepOnePlus Git 30 7300 31 7500 34 http www takara bio coJp 8 RR820A lt Applied Biosystems 7500 Fast Real Time PCR System gt PCR Holding Stage Reps 1 95 C 30 Cycling Stage Number of Cycles 40 95 C 3 60 3
22. 100 ng 2 2 ul dH2O 8 5 ul Total 25 ul 3 1 primer 0 4 uM 0 2 1 0 UM 2 template DNA template 100 ng RT PCR cDNA RT template PCR 10 3 25 ul 2 PCR PCR PCR Tm PCR 3 PCR PCR Panem ZEE Hold 1 107 Cycle 1 95 C 30 ei 2 Step PCR 4 Cycle 40 95 C 5 60 30 60 Hold Tie deg 5 5 t me mm as 00 05 Data Colecon aj
23. ll 200 50 ul SYBR Premix Ex Taq Il Tli RNaseH Plus 2conc 1 1mlX5 ROX Reference Dye 50 X conc 2 200 ul ROX Reference Dye Il 50 Xconc 2 200 ul x 1 TaKaRa Ex Taq HS dNTP Mixture Mg2 Tli RNase H SYBR Green 16 2 Life Technologies PCR ROX Reference Dye Applied Biosystems 7300 Real Time PCR System Life Technologies StepOnePlus Real Time PCR System Life Technologies ROX Reference Dye Il Applied Biosystems 7500 7500 Fast Real Time PCR System Life Technologies Thermal Cycler Dice Real Time System TP900 TP960 LightCycler LightCycler 480 Roche Diagnostics CFX96 Real Time PCR Detection System Bio Rad 1 PCR authorized instruments 2 3 PCR 4 5 x
24. ug ADNA 74 1 DNA http wwwr takara bio coJp 16 RR820A IX SYBR Premix Ex Taq Il Tli RNaseH Plus RR820S B L W LR WR SYBR Premix Ex Taq Tli RNaseH Plus RR420S A B L W LR WR SYBR Premix DimerEraser Perfect Real Time RRO91A B SYBR Premix Ex Taq GC Perfect Real Time RRO71A B PrimeScript RT reagent Kit Perfect Real Time RR037A B PrimeScript RT Master Mix Perfect Real Time RR036A B PrimeScript RT reagent Kit with gDNA Eraser Perfect Real Time RR047A B One Step SYBR PrimeScript RT PCR Kit Perfect Real Time RRO66A B One Step SYBR PrimeScript RT PCR Kit Il Perfect Real Time RRO86A B One Step SYBR PrimeScript PLUS RT PCR Kit Perfect Real Time RRO96A B Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice Real Time System Single TP850 TP870 Thermal Cycler Dice Real Time System Lite TP700 TP760 Perfect Real Time http wwwr takara bio coJp realtime
25. 0 Melt Curve Stage Takaka Ex Taq HS 7ag PCR PCR PCR 95C 5 15 PCR 95 30 3 http wwwr takara bio coJp 9 RR820A lt LightCycler LightCycler 480 gt X Roche Diagnostics PCR lt 1 gt SYBR Premix Ex Taq I Tli RNaseH Plus 2X 10 ul 1X PCR Forward Primer 10 uM 0 8 ul 0 4 uM PCR Reverse Primer 10 uM 0
26. 8 ul 0 4 uM template lt 100 ng 2 2 ul dH2O 6 4 ul Total 20 ul 1 primer 0 4 UM 0 2 1 0 UM 2 template DNA template 100 ng RT PCR cDNA RT template PCR 10 2 PCR PCR PCR Tm PCR 3 PCR PCR 12 lt LightCycler gt PCR Stage 1 onpas Tiaaton Rabe C 00 95 C 30 20 C ToyeTerewowv C
27. O 1 Mire Delay zycles a Stage 2 PCR AR Jf 95 C 5 20 60C 20 20 40 Stage 3 95 0 20 65 15 20 95 0 0 1 http www takara bio co jp 10 RR820A lt LightCycler 480 gt cA PUR me TaS Tee oD EE IL Ramo Ree CN AmhNin wm OQ Sm Tage Q Sup See OQ Seep Delay Gorden br oe s aj DE gjere Foo 03 0 iss a o FE H Rere 00109296 RI a FO PCR Denature 95 C 30 Ramp Rate 4 4 C sec 1 PCR Analysis Mode Quantification 95 C 5 Ramp Rate 4 4 C sec 60 C 30 Ramp Rate 2 2 C sec Acquisition Mode Single 40 Melting Analysis Mode Melting Curves 95 C 5 Ramp Rate 4 4 C sec 60 1 Ramp Rate 2 2 C sec 95 Ramp Rate 0 11 C sec Acquisition Mode Continuous Acquisitions 5 per 1 Cooling 50 C 30 Ramp Rate 2 2 C sec 1 TaKaRa Ex Taq H5 7gg PCR PCR
28. claims 6 16 outside the U S corresponding to the expired U S Patent No 5 436 149 M82 Tli RNaseH Plus This product is the subject of the pending JP patent application http wwwr takara bio coJp 18 RR820A http www takara bio co jp 19 RR820A TafdR4 Tel 077 543 6116 Fax 077 543 1977 http www takara bio co jp SII AA RNS AL v201308Da
29. earch Foundation or Evotec Biosystems GmbH and licensed to Idaho Technology Inc and Roche Diagnostics GmbH to use only the enclosed amount of product according to the specified protocols No right is conveyed expressly by implication or by estoppel to use any instrument or system under any claim of U S Patent Nos 6 174 670 6 569 627 and 5 871 908 other than for the amount of product contained herein L52 Rox Reference Dye Research Field Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US 5 928 907 The purchase of this product includes a limited non transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser s own internal research No right under any other patent claim and no right to perform commercial services of any kind including without limitation reporting the results of purchaser s activities for a fee or other commercial consideration is conveyed expressly by implication or by estoppel This product is for research use only Further information on purchasing licenses may be obtained by contacting the Director of Licensing Applied Biosystems 850 Lincoln Centre Drive Foster City California 94404 USA 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 M57 LA Technology This product is covered by the
30. mer 10 uM 1 ul 0 4 uM 1 PCR Reverse Primer 10 uM 1 ul 0 4 uM 1 template lt 100 ng 2 2 ul dH2O 8 5 ul Total 25 ul 3 1 primer 0 4 uM 0 2 1 0 UM 2 template DNA template 100 ng RT PCR cDNA RT template PCR 10 3 25 ul 2 PCR PCR PCR PCR Panem Mas lt u Hold 107 Cycle 1 VY 95 C 30 2 Step PCR 4 Cycle 40 95 C 5 60 C 30 old Tin mm ss 0 Dissociation Ota Cofecton 4 PCR 12
31. waar RR820A IdfaR4 SYBR Premix Ex Taq Il Tli RNaseH Plus v201308Da SYBR Premix Ex Taq Tli RNaseH Plus t SYBR Greenl PCR 2 SYBR Green RNase H Tli RNase H 2 cDNA mRNA PCR SYBR Premix Ex Taq Tli RNaseH Plus RR420A 7gg PCR ARR TaKaRa Ex Tag HS PCR
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