Mouse Feeder Cell Quantification Kit
Contents
1. 7 PCR PCR Thermal Cycler Dice Real Time System VII Appendix Hold Cycle 1 95 30 2 step PCR Cycle 40 95 5 60 30 FAM 8 http wwwr takara bio coJp 10 RR290 mtDNA FAM nDNA mtDNA Ct nDNA mtDNA 1 nDNA nDNA nDNA 6 02 x 1023 Xx g9 bp2. 3 DNA 100 DNA 3 hES 1 X 109 DNA DNA a b pg Ct pg Ct 200 pg 21 62 20 000 pg 24 43 20 pg 25 21 4 000 pg 26 69 2 pg 28 88 800 pg 29 16 0 2 pg 2 160 pg 31 45 Y 3 718 X 30 103 Y 3 366 X 38 882 R2 0 9996 R2 0 9997 c Ct DNA DNA DNA 5 Ct pg Ct pg pg 19 51 24 32 21 168 21 875 3 230 22 57 24 21 22 822 22 932 0 476
3. A DNA Target List STD pg 100pg 2 ul 200 po B mtDNA Target List 1 STD 3 Hold Cycle 1 95 30 2 step PCR Cycle 40 95 5 60 30 FAM 4 Start Run Run File http wwwr takara bio coJp 8 RR290 5 FAM Result Analysis Analysis Data Standard Curve Fluorescence 2nd Derivative Max R2 0 97 Flunnescertee nd emmahm Ma 1
4. mtDNA DNA mtDNA 2 A DNA DNA PCR DNA DNA Ct pg pg DNA DNA
5. Ct 35 2 0 97 0 97 http wwwr takara bio coJp 14 RR290 IX DNA DNA mtDNA Ct Ct DNA DNA
6. CNV 3 primer 1 Type1 3 Type ll 7 Type 15 Thermal Cycler Dice Real Time System 1 SYBR Premix Ex Tag ll Ti RNaseH Plus 2 Xx conc 12 5 ul Mouse_Mt_primer 10 uM each 1 0 wl dH20 9 5 ul Total 23 ul 1 SYBR Premix Ex Tag 1 Ti RNaseH Plus 2 Xx conc 12 5 ul Human_primer 10 uM each 1 0 ul dH2O 9 5 ul Total 23 http wwwr takara bio coJp 6 RR290 Life Technologies PCR 1 SYBR Premix Ex Tag 1 Ti RNaseH Plus 2 Xx conc 12 5 ul Mouse_Mt_primer 10 uM each 1 0 ul ROX Reference Dye or ROX Reference Dye ll 3 0 5 ul dH2O 9 0 ul Total 23 bl 1 SYBR Premix Ex Tag 1 Tli RNaseH Plus 2 Xx conc 2 S50 Human_primer 10 uM each 1 0 ul ROX Reference Dye or ROX Reference Dye II 3 0 5 ul dH2O 9 0 ul Total 23 ul 3 StepOnePlus ROX Reference Dye Applied Biosystems 7500 Fast Real Time PCR System ROX Reference Dye I 4
7. Thermal Cycler Dice SYBR Molecular Probes Inc Premix 797 http wwwr takara bio coJp 15 RR290 sm Iaf4R4 Tel 077 543 6116 Fax 077 543 1977 http www takara bio co jp v201407Da
8. 10 ng ul 1ng ul 100 pg ul 10 pg ul 10 pg ul Thermal Cycler Dice Real Time System nDNA 1 SYBR Premix Ex Tag Il Tli RNaseH Plus 12 5 4 Mouse_genome_primer 10 uM 1 0 ul dH20 9 5 ul Total 23 mtDNA 1 SYBR Premix Ex Tag Il Tli RNaseH Plus 12 5 Mouse_Mt_primer 10 uM 1 0 ul dH20 9 5 ul Total 23 ul http www takara bio co jp 9 RR290 Life Technologies PCR nDNA 1 SYBR Premix Ex Tag Il Tli RNaseH Plus S Mouse_genome_primer 10 uM 1 0 wl ROX Reference Dye or ROX Reference Dye II 3 0 5 ul dH20 9 0 ul Total 23 mtDNA 1 SYBR Premix Ex Tag Il Tli RNaseH Plus 12 5 Mouse_Mt_primer 10 uM 1 0 ul ROX Reference Dye or ROX Reference Dye II 3 0 5 ul dH2O 9 0 ul Total 23 3 StepOnePlus ROX Reference Dye Applied Biosystems 7500 Fast Real Time PCR System ROX Reference Dye Il 4 0 2 ml 1 23 ul 5 2 6
9. DNA http wwwr takara bio coJp 2 RR290 B DNA mtDNA PCR mtDNA DNA nDNA DNA nDNA mtDNA PCR nDNA Ct SDM mtDNA Ct Ct nDNA mtDNA DNA 1 PCR E 100 25 IPCR 1 2 3 4 5 6 7 8 9 10 11 SYBR Premix Ex Tag 1l Tli RNaseH Plus 2 Xx Conc 625 UIX 4 Mouse_Mt_primer 10 uM each 100 wl Mouse_genome_ primer 10 uM each 100 ul Human_primer_Type 1 CHR3 10 uM each 100 ul
10. bp 3 3 x 1032 660 nDNA 20 000 pg 5 528 X 103 2 000 pg 5 528 X 102 200 pg 5 528 Xx 101 20 pg 5 5 6 nDNA 2 nDNA nDNA Ct 3 mtDNA Ct 2 mtDNA 4 mtDNA nDNA nDNA mtDNA mtDNA mtDNA nDNA mtDNA nDNA mtDNA hES MEF 1 x 10 DNA nDNA DNA a nDNA pg nDNA Ct 20 000 pg 5 528 X 103 24 64 2 000 pg 5 528 Xx 102 27 81 200 pg 5 528 Xx 10 30 36 20 pg 5 526 33 93 Y 3 044 X 36 016 R2 0 9965
11. PCR DNA SYBR Premix Ex Tag 11 Tli RNaseH Plus RR820S A B NucleoSpin Tissue 740952 10 50 250 NucleoSpin Tissue XS 740901 10 50 250 Human Genomic DNA 636401 Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice Real Time System Lite TP700 TP760 Thermal Cycler Dice Real Time System Single TP850 TP870 0 2 ml Hi 8 Tube NJ300 0 2 ml Hi 8 Dome Cap NJ301 96 well Hi Plate for Real Time NJ400 Sealing Film for Real Time NJ500 XI
12. 20 IV 1 DNA VI A 2 DNA NucleoSpin Tissue 740952 10 50 250 NucleoSpin Tissue XS 740901 10 50 250 3 PCR 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 4 5 PCR 0 2 ml Hi 8 Tube NJ300 0 2 ml Hi 8 Dome Cap NJ301 96 well Hi Plate for Real Time NJ400 Sealing Film for Real Time NJ500 6 V V 1 mtDNA Symbol COXI_ MM IMU Rplp1 9 NMO18853 Human_primer_Typel CHR3 ALAS1 NM_000688 Human_primer_Typell CHR7 PPIA NM_021130 Huma
13. 0 2 ml 1 23 ul 5 2 6 7 PCR PCR Thermal Cycler Dice Real Time System VII Appendix Hold Cycle 1 95 30 2 step PCR Cycle 40 95 5 60 30 FAM 8 http wwwr takara bio coJp 7 RR290 DNA FAM DNA 1 2
14. RR290 IdRaRa Mouse Feeder Cell Quantification Kit v201407Da 1 hESC hiPSC A Mouse Feeder Cell Quantifcation Kit 16 hiPSC DNA i DNA PCR DNA mt DNA 4 PCR mtDNA
15. http wwwr takara bio coJp 1 1 RR290 b mtDNA Ct mtDNA 25 40 3074 5 528 hES MEF nDNA 556 DNA mtDNA VIl Appendix Thermal Cycler Dice Real Time System Thermal Cycler Dice Real Time System 1 AQ S Absolute Quantifcation Single 2 Plate Setup Target amp Sample Setting Dye FAM Sample List STD UNKN Update Tasget et Target File Templale Load fd _ Ag Dm Ip lpye JINeme caioe IXwr fet iAsmwyD Forward Forward Tm Reverae ReveseTm Prodoclsize ima FRM Human am nose mm http www takara bio co jp 12 RR290 Plate Setup Plate Image Update
16. 998 3 _SetsTp oe TB LO i Result Analysis Analysis Data Text Report Fluores cence Data Set of Each Well Data Set of Replicate A DNA OQty SDM DNA X 100 B mtDNA Qty SDM mtDNA nDNA mtDNA mtDNA mtDNA nDNA VIII 1 Mu 9 Tm
17. 100 pg ul 10 pg uk DNA 1 1 pg u 0 1 pg ul hES hiPS DNA 10 ng ul 2 ng u DNA Human Genomic 400 pg uu 80 pg ul DNA 2 1 DNA mtDNA mtDNA DNA 2 hES hiPS Human Ge nomic DNA DNA 1 1 X 106 1 X 107 DNA 3 3 NucleoSpin Tissue NucleoSpin Tissue XS DNA http wwwr takara bio
18. Human_primer_Type ll CHR7 10 uM each 100 Human_primer_Type lll CHR15 10 uM each 100 dH2O 1 mlX 2 EASY Dilution for Real Time PCR 1 mlx 2 ROX Reference Dye 50 Xx Conc 50 ul ROX Reference Dye ll 50 Xx Conc 50 Human Genomic DNA 100 ng ul 20 ul Life Technologies PCR ROX Reference Dye StepOnePlus Real Time PCR System Life Technologies ROX Reference Dye I Applied Biosystems 7500 Fast Real Time PCR System Life Technologies Thermal Cycler Dice Real Time System TP900 TP960 http wwwr takara bio coJp 9 RR290 20 80 SYBR Premix Ex Tag ll Tli RNaseH Plus 4 6 80 20 4 6
19. coJp 5 RR290 2 DNA a DNA 10 ng ul EASY Dilution b DNA EASY Dilution DNA 100 pg ul 10 pg u 1 pg ul 0 1 pg ul 10 DNA 10 ng u 2ng u 400 pg u 80 pg ul 5 3 PCR qa 1 mtDNA DNA Human_primer_Type rimer M Mt_ primer 9 DI 1 2 DNA DNA DNA 4 4 DNA 10 ng ul 2 ul 1 Primer mix 2 Human_primer CNV 3
20. DNA A 3 23 B 0 478 C 0 059 http wwwr takara bio coJp 8 RR290 B DNA mtDNA 1 DNA 1 NucleoSpin Tissue NucleoSpin Tissue XS DNA 2 EASY Dilution DNA nDNA 10 ng l 1ng 100 pg ul 10 pg 10 DNA mtDNA 10 pg ul 3 PCR nDNA mtDNA o 2 2 primer mix EE ET TT primer Mouse_genome_ primer Mouse_Mt_primer DNA
21. n_primer_Type Ill CHR15 NM_004048 1 M mitochondria http wwwr takara bio coJp 4 RR290 V 2 A DNA mtDNA DNA Mouse_Mt_primer Human_primer_Typel 1 2 2 Human_primer CNV 3 CNV Type1 3 Type ll 7 Type 15 primer 1 B mtDNA DNA lt 2DNAN Mouse_genome_primer Mouse Mt primer VI A DNA DNA OO DNA DNA DNA
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