RR041A_j
Contents
1. SYBR Molecular Probe Inc http wwwr takara bio coJp 15 RR041A NOTICE TO PURCHASER LIMITED LICENSE P5 PCR Notice Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US 5 994 056 and 6 171 785 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 ex2. O No Template Control O Ct PCR 100 0 1 uM 1 0 uM PCR O
3. SYBR Premix Ex Taq 2Xconc 80 SYBR Green http www takara bio co jp 4 RR041A VI lt Thermal Cycler Dice Real Time System gt 1 PCR
4. PCR 95 C 5 95 5 60 30 64 C O 3 step PCR X PCR 3 te PCR p gt 9 o 7 95 C 5 95 C 5 D 5 A we 5 P 60 C 60 1 55 C 30 556 726 3 726 1 95 30 DNA 95 C 2 2 http www takara bio co jp 11 RR041A
5. RT PCR amp Perfect Real Time http www takara bio co jp realtime RefSeq lt RT PCR SYBR Green RT PCR RT PCR PCR P 5 10 80 150 bp 300 bp 40 60 45 55 Tm Forward primer Reverse primer Tm Tm OLIGO 1 63 68 C Primer3 2 60 65 C
6. c PCR 95 C 30 3 PCR http www takara bio cojp 9 RRO41A gt Mx3000P gt Mx3000P Stratagene 1 PCR gt 1 gt SYBR Premix Ex Taq 2 X 12 5 ul 1x PCR Forward Primer 10 uM 0 5 ul 0 2 uM 1 PCR Reverse Primer 10 uM 05 ul 0 2 uM 1 ROX Reference Dye 50x 2 05 ul 1x template lt 100 ng 2 ul 3 dH2O 9 ul Total 25 ul 1 primer 0 2 UM 0 1 1 0 uM 2 ROX Referenc
7. SYBR Green Assay PCR 3 SYBR Premix Ex Taq Perfect RealTime RR041 SYBRS Premix Ex Taq Perfect Real Time RRO81 SYBR Premix Ex Taq Tli RNaseH Plus RR820 SYBR Premix DimerEraser Perfect Real Time RR091 SYBR Premix Ex Taq SYBR Premix Ex Taq SYBR Premix DimerEraser w VII Appendix 1 PCR
8. PCR MC Hold Cycle 1 95 C 30 2 Step PCR Cycle 40 95 C 5 60 C 30 Dissociation TaKaRa Ex Taq HS 7ag PCR PCR PCR BID 95 C 5 15 PCR 95 30 3 Thermal Cycler Dice Real Time System Thermal Cycler Dice Real Time System http www takara bio co jp 5 RR041A lt ABI PRISM 7000 7700 Applied Biosystems 7300 7500 7500 Fast Real Time PCR System
9. 4 80 20 4 6 http wwwr takara bio coJp 3 RR041A IV 1 2 3 V PCR SYBR Green 2 X conc PCR PCR Hot Start PCR FARES TaKaRa Ex Tag HS PCR 1 2 3 4
10. 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 http wwwr takara bio coJp 12 RR041A 1 OLIGO Primer Analysis Software Molecular Biology Insights 2 Primer3 http www genome wi mit edu ftp distribution software 3 http www ncbinlIm nih goVBLAST Perfect Real Time TEL 03 3271 8553
11. SYBR Molecular Probes Inc 2 Thermal Cycler Dice Real Time System TP900 TP960 SYBR Premix Ex Taq 1 RR820A B RR081A B 3 Smart Cycler Cepheid 7aKaRa Ex Taq Hs PCR PCR SYBR Greenl 1 PCR PCR DNA DNA DNA 3 1 100 Hot Start PCR TaKaRa Ex Taq HS 28 D
12. 7ag PCR PCR PCR 95 C 5 15 PCR 95 30 3 PCR http wwwr takara bio coJp 10 RR041A PCR PCR Perfect Real Time PCR RR820 RR081 RR091
13. Tm PCR 3 PCR PCR 11 Cycle Program Data yelesl 40 5 Analysis Mode None Melting Curves Temperature Targets Target Temperature C Incubation Time hrs min sec Temperature Transition Rate C sec Wa Temperature C Step Size C Step Delay cycles Acquisition Mode 5 a Gan 9 Geis 55 koh 56 e po poo ph goo Nn Cm Stage 2 PCR PCR Stage 1 956 30 1 Stage 2 PCR 95 5 606 20 20 40 Stage 3 95 0 20 65 15 20 95 C OF TaKaRa Ex Taq HS 7a9 PCR PCR PCR 95 5 15
14. TEL 077 543 7297 2 RT PCR RT PrimeScript RT RR037A B RR036A B RR047A B PrimeScript RT reagent Kit Perfect RealTime F RR037A 1 RNA o RNA lt 1 gt 5 X PrimeScript Buffer for Real Time 2 ul 1 PrimeScript RT Enzyme Mix 0 5 ul Oligo dT Primer 50 uM 1 05 ul 25 pmol Random 6 mers 100 uM 05 ul 50 pmol total RNA RNase Free dH2O Total 10 ul 2 1 Oligo dT Primer Random 6 mers mRNA cDNA Gene Specific Primer Oligo dT Primer 50 uM 0 5 ul 25 pmol Random 6 mers 100 uM 0 5 ul 50 pmol
15. gt LightCycler Roche Diagnostics 1 PCR gt 1 gt SYBR Premix Ex Taq 2 X PCR Forward Primer 10 uM PCR Reverse Primer 10 uM template lt 100 ng dH2O Total 10 ul 1x 04 ul 0 2 uM 1 0 4 ul 0 2 uM 1 2 ul 2 7 2 ul 20 ul 1 primer 0 2 UM 0 1 1 0 uM 2 template DNA template 100 ng RT PCR cDNA RT template PCR 10 2 PCR LightCyclery PCR PCR PCR
16. 12 5 ul 1x 05 ul 02 uM 1 05 ul 02 uM 1 2 ul 2 95 ul 25 ul 1 primer 0 2 UM 0 1 1 0 UM 2 template DNA template 100 ng RT PCR cDNA RT template PCR 10 2 Smart Cycler Smart Cycler PCR PCR PCR Tm PCR 3 PCR 11 PCR Sane2 Re
17. ABI PRISM 7000 7700 4 7300 Real Time PCR System ROX Reference Dye 50 xX 4 http wwwr takara bio coJp 6 RR041A 2 PCR PCR PCR Tm PCR 3 PCR 11 gt ABI PRISM 7000 7700 7300 7500 Real Time PCR System gt Thermol Cycler Protocol PCR Themel Profile Auto Increman Ramp Rate stage1 Stage 2 Stage 3 Stage 1 Reps 1 Reps Reps 1_ Reps 1 95 C 30 Stage 2 PCR Reps 40 95 C 5 60 C 30 34 Stage 3 Melt Curve 7700 30 7000 7300 31 7500 34 gt 7500 Fast Real Time PCR System gt PCR Holding Stage
18. gt 1 PCR gt 1 gt SYBR Premix Ex Taq 2 X 10 ul 25 ul 1 X PCR Forward Primer 10 uM 0 4 ul 1 ul 0 2 uM 1 PCR Reverse Primer 10 uM 0 4 ul 1 ul 0 2 MM 1 ROX Reference Dye 50 X or Dye ll 50 x 3 04 ul 1 ul 1x template 2 ul 4 ul 2 dH2O 6 8 ul 18 ul Total 20 pl 4 50 ul 4 1 primer 0 2 UM 0 1 1 0 UM 2 template 20 ul DNA template 100 ng RT PCR cDNA RT template PCR 10 3 ROX Reference Dye Il 50X ROX Reference Dye 50 7500 Real Time PCR System 7500 Fast Real Time PCR System ROX Reference Dye Il 50x
19. gt 1 gt SYBR Premix Ex Taq 2 X 12 5 ul 1x PCR Forward Primer 10 uM 05 ul 0 2 uM 1 PCR Reverse Primer 10 uM 05 ul 0 2 uM 1 template lt 100 ng 2 ul 2 dH2O 9 5 ul Total 25 ul 3 1 primer 0 2 uM 0 1 1 0 uM 2 template DNA template 100 ng RT PCR cDNA RT template PCR 10 3 25 pl 2 PCR PCR PCR Tm PCR 3 PCR 11
20. Fluorescence Cycles Fluorescence 21 60 70 80 90 Degrees C RT PCR Human GAPDH mRNA total RNA 1 pg 100 ng cDNA Negative Control dH20 http www takara bio co jp 14 RR041A VII SYBR Premix Ex Tag 5 mL 25 mL RRO41L W SYBR Premix Ex Tag 5 mL 25 mL ROX plus RR41LR WR SYBR Premix Ex Tag Tli RNaseH Plus RR420A B SYBR Premix Ex Taq II Tli RNaseH Plus RR820A B SYBR Premix Ex Tag Perfect Real Time RRO81A B SYBR Premix Ex Tag 11 5 mL 25 mL RRO81L W SYBR Premix Ex Tag 11 5 mL 25 mL ROX plus RR81LR WR SYBR Premix DimerEraser Perfect Real Time RR091A 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 RRO47A B One Step SYBR PrimeScript PLUS RT PCR Kit Perfect Real Time RR096A B One Step SYBR PrimeScript RT PCR Kit Perfect Real Time RRO86A B One Step SYBR PrimeScript RT PCR Kit Perfect Real Ti
21. 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 Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser s own internal research No other patent rights are conveyed expressly by implication or by estoppel 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 M57 LA Technology This product is covered by the claims 6 16 of U S Patent No 5 436 149 and its foreign counterpart patent claims Takaka Tel 077 543 6116 Fax 077 543 1977 http www takara bio co jp SINT ANS 4l 201108Da
22. Reps 1 95 C 30 Cycling Stage Number of Cycles 40 95 C 3 60 30 Melt Curve Stage TaKaRa Ex Taq HS 7gg PCR PCR PCR 95 5 15 PCR 95 30 3 PCR http www takara bio coJp 7 RR041A gt Smart Cycler System gt 1 PCR lt 1 gt SYBR Premix Ex Taq 2 X PCR Forward Primer 10 uM PCR Reverse Primer 10 uM template lt 100 ng dH2O Total
23. Gene Specific Primer 2 UM 0 5 ul 1 pmol K2 10 ul 500 ng total RNA 2 37 15 3 85 5 4 C 3 Gene Specific Primer 42 15 PCR 50 http wwwr takara bio coJp 13 RR041A 3 PCR Smart Cycler System o 22 5 24 ul gt 1 gt SYBR Premix Ex Taq 2 X PCR Forward Primer 10 uM PCR Reverse Primer 10 uM dH20 Total 12 5 ul 1x 05 ul 0 2 uM 05 ul 0 2 uM ul 22 5 24 ul 4 1 25 ul PCR 2 5 150
24. NA SYBR Green KE DNA DNA 1 F F OMA F F F 1 1 1 1 1 E 2 F F eM F F F 3 a gt F PX 4 OE do a http www takara bio co jp 2 RR041A ll AB 200 50 ul SYBR Premix Ex Taq Perfect Real Time 2 X conc 1 1mlx5 ROX Reference Dye 50 X conc 2 200 ul ROX Reference Dye Il 50 X conc 2 200 ul 1 TaKaRa Ex Taq HS dNTP Mixture Mg2 amp S amp U SYBR Green 2 Life Technologies PCR ABI PRISM 7000 7700 7300 Real Time PCR System ROX Referenc
25. e Dye 7500 Real Time PCR System ad 7500 Fast Real Time PCR System ROX Reference Dye ROX Reference Dye Thermal Cycler Dice Real Time System Smart Cycler System LightCycler SEBUM CBE PCR authorized instruments PCR Sia el PCR VII 0 RT PCR amp Perfect Real Time http wwwr takara bio coJp realtime Ill 4 6
26. e Dye Il 50x ROX Refer ence Dye 50 X Mx3000P 3 template DNA template 100 ng RT PCR cDNA RT template PCR 10 2 PCR PCR PCR Tm PCR 3 PCR PCR 11 PCR Segment 1 95 C 1 Segment 2 PCR 95 C 5 60 20 40 Segment 3 Dissociation Curve TaKaRa Ex Taq HS
27. me RRO66A Premix Ex Tag Probe qPCR RR390A B One Step PrimeScript RT PCR Kit Perfect Real Time RR064A Smart Cycler System SC200N SC210N Smart Cycler Software Version 2 0 Upgrade Kit SC220 Perfect Real Time http www takara bio co jp realtime RefSeg lt RT PCR SYBRS Green RT PCR 1 2005 3 4 PCR 120 126 2 2008 PCR 3 1 RT PCR 39 43
28. peat 40 times 2 Tempos ature Cycle v Stage 1 Hold v _Optcs on Temp Secs 95 0 3 Na 950 15 NA 600 20 On DegiSec Temp Secs Optics on Stage 3 Stage 1 Molt Curve X Hold 95 30 Stage 2 PCR Repeat 40 times 95 C 5 60 20 Stage 3 Melt Curve TaKaRa Ex Taq H5 7a9 PCR PCR PCR 95 5 15 PCR 95 C 30 3 Smart Cycler System Smart Cycler System VII http www takara bio co jp 8 RR041A gt LightCycler
29. pressly by implication or by estoppel This product is for research use only Diagnostic uses under Roche patents require a separate license from Roche 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 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 patent 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 providing 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 veterinary 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 Research 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
30. waar RRO41A IdfaR4 SYBR Premix Ex Taq Perfect Real Time v201108Da SYBR Premix Ex Taq Perfect Real Time SYBR Green 1 PCR 2 X SYBR Green PCR TaKaRa Ex Taq HS PCR PCR Thermal Cycler Dice Real Time System TP800 TP850 2 Smart Cycler System 3 Smart Cycler System 3 Cepheid ABI PRISM 7000 7700 Applied Biosystems 7300 7500 Real Time PCR System Applied Biosystems 7500 Fast Real Time PCR System Life Technologies LightCycler Roche Diagnostics Mx3000P Agilent Technologies 1 Molecular Probes Inc SYBR Green 1l
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