PRIMARY CELL
Contents
1. 5 lt gt TaqMan gt Applied Biosystems 7900HT PCR lt gt SYBR ExScript RT PCR Kit Perfect Real Time TAKARA RR053A lt gt2. PAI 1 HPRT 5 o 4 3 2 1 0 0 1 2 3 4 5 6 7 8 Days Fig 1 PAI 1 Adiponectin HPRT 400 A 300 3 200 100 0 0 1 2 3 4 5 6 7 8 Days Hig 2 Adiponectin Leptin HPRT 400 300 200 IO Rat 100 Fig 3 Leptin Insulin Receptor HPRT N wo A Ratio Days Fig 4 Insulin Receptor PPAR y HPRT Fig 5 PPARy lt gt 100 Fig 6 7 cDNA Fig T2 0 997 E Real time PCR dissociation curve Fig 8
3. 1 000 Er1 Amplification Plot 1 000 1 000 E 1 amp Zi 000 E2 1000 E3 1 000 E 4 i 1 000 E 5 Fig 6 36 Standard Plot Legend a m Standards 32 Unknowns X Show Unknowns 28 Standard Curve 24 22 Slope 3 3060367 20 Y Inter 37 05646 18 R2 0 9978927 16 14 12 1 0 1 0 EH 1062 10E 1064 1065 1066 106 Quantity Fig 7 1 Standard Plot ka 32 m Standards 30 x Unknowns 28 Hide Unknowns 26 EE 5 24 Standard Curve 22 Slope 3 2414346 20 Y Inter 34 29514 18 R2 0 99703544 16 14 12 1 0 1 0 EH 10 E 2 10 E33 1064 1 0 EH 1 0 E46 Quantity Fig 7 2 x Dissociation Curve 2 400 E1 1 900 E1 1 400 E1 Derivative 9 000 E2 4 000 E2 MEE mm mm 65 0 70 0 75 0 80 0 85 0 90 0 95 0 E Temperature C Fig 8 lt gt puces VAC01 V1 C7 v b BOR BAT01 E BATO2 8 LE WATO1 1 WATO2 8 Fy by M AKO1 Ele a AK
4. PAL1 100 1 20 C 100 w 1 Adiponectin 100 1 20 C 10041 Leptin 100 1 20 C 100 41 Insulin 100 1 20 C Receptor 100 w 1 PPAR y 100 1 20 C 10041 HPRT 100 1 200 10041 PAT 1 108 1 1 20 C 50 w 1 Adiponectin 108 1 1 20 C 501 Leptin 108 1 20 50z1 Insulin 108 1 1 20 C Receptor KJ 50u41 PPAR y 108 1 1 20 C 50 w 1 HPRT 108 1 20 C 501 PAT 1 PPAR y HPRT Plasminogen Activator Inhibitor 1 Peroxisome Proliferator Aactivated Receptor gamma Hypoxanthine Phosphoribosyltransferase 1 BU gt 1
5. PCR Code PCRO1 PRIMARY CHL D PCR 18 2 PAI 1
6. 1027 106 2 HPRT HPRT Vs 384 2041327 3500rpm for 1min 1 1 95 C 10sec 2 40 95C 5sec 60 C 30sec gt VAC Visceral Adipocyte Culture Kit 0 8 RNA HPRT 0 1 Hig 1 5
7. SYBR RT PCR Kit Perfect Real Time Takara Code RR053A C PCR PCR 5 X M MLV buffer 4ul dNTP Mixture 10mM each lul Random 6mers 100 u M lul Oligo dT primer 50 u M lul M MLV RTase 200U u 1 0 5 ul RNase Inhibitor 40U u 1 0 5ul total RNA lug RNase Free dH20 Total volume 20 ul 42C 15min 95 C 2min 2 PCR SYBR RT PCR Kit Perfect Real Time Takara Code RR053A Applied Biosystems 7900HT PCR PCR 1 SYBR Premix Ex TaqTM 2x 10 1 PCR primer Mixture 10 MD 0 8 1 ROX Reference Dye 50 x 0 41 template cDNA lul RNase Free dH20 7 8 1 Total volume 20 1 1 3 cDNA PCR
8. O06 DNA AKO09 http www primarycell com gt PCR http www gene lab com DNA Affymetrix t Gene Chip http www gene lab com 10 T061 3242 3 761 2 TEL 0133 60 2880 FAX 0133 60 2881 tech primarycell com 060 0009 9 15 28 196 IT 3F TEL 011 644 7333 FAX 011 644 7611 adv support gene lab com 11
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