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05-8 SYBR Green Realtime PCR Master Mix (Code No. QPK-201, QPK-201T) TOYOBO CO., LTD. Life Science Department OSAKA JAPAN A3314K

(2) (2) (3) (3) (5) ABI PRISM 7700 (5) Roche LightCycler TM (7) BioFlux LineGene (9) 1 RT-PCR (10) (12) (13) F. Hoffmann-La Roche Ltd. Roche Molecular Systems Inc. The Perkin-Elmer Corporation Purchase of this product is accompanied by a limited license to use it in the Polymerase Chain Reaction (PCR) process for The Research Field in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by the up-front license fee, either by payment to Perkin-Elmer or as purchased, i.e., an authorized thermal cycler. LightCycler TM Idaho Technology Inc. Roche Molecular Systems Inc. SYBR Molecular Probes Inc. ABI PRISM The Perkin-Elmer Corporation - - 1 - -

DNA 2 SYBR Green I(Molecular Probes Inc.) PCR LightCycler TM BSA ABI PRISM 7700 LightCycler TM LineGene Taq DNA Taq PCR 10 1 LightCycler TM PCR -20 2 4 (QPK-201 1-20 4 ) 1 4 2 ( ) -20 4 80 20 - - 2 - -

[SYBR Green Realtime PCR Master Mix] dntp MgCl 2 Taq DNA Taq PCR SYBR Green I 2 [ ] PCR 20 30mer GC 40 60% 200bp 150bp 50 150bp [ ] PCR [ PCR ] ABI PRISM 7700 LightCycler TM LineGene PCR - - 3 - -

DNA [cdna] 1st strand cdna 1st strand cdna dt PCR 1st strand cdna ReverTra Ace - - (Code:FSK-101) 10 cdna ( ) PCR [ DNA ] PCR DNA 1 10ng DNA - - 4 - -

ABI PRISM 7700 (1) [PCR ( )]( 0.4 M 50 L) 16 L SYBR Green Realtime PCR Master Mix 25 L 1 (10 M) 2 L 2 (10 M) 2 L 5 L 50 L SYBR Green Realtime PCR Master Mix 1/2 ( ) 0.2 0.6 M (2)PCR 3 ( 1) Tm 55 65 60 15 PCR 1 5 Detector SYBR Quencher None PCR data collection 30 2 ( 2) data collection - - 5 - -

[PCR ( 1)](3 60 72 ) 95 60 95 15 40 60 15 72 45 (data collection) [PCR ( 2)](2 60 ) 95 60 95 15 40 60 60 (data collection) - - 6 - -

Roche LightCycler TM (1) [PCR ( )]( 0.4 M 20 L) 6.4 L SYBR Green Realtime PCR Master Mix 10 L 1 (10 M) 0.8 L 2 (10 M) 0.8 L 2 L 20 L SYBR Green Realtime PCR Master Mix 1/2 ( ) 0.2 0.6 M (2)PCR Tm 55 65 55 10 20 5 30 5 PCR 1 5 200bp 15 data collection 10 ( ) Detector F1 Temperature Transition Rate 20 /sec( ) 2 data collection - - 7 - -

[PCR ( )](3 55 72 ) 95 30 95 5 40 55 10 72 15 (data collection) (Melting Curve) - - 8 - -

BioFlux LineGene (1) [PCR ( )]( 0.4 M 10 L) 2.2 L SYBR Green Realtime PCR Master Mix 5 L 1 (10 M) 0.4 L 2 (10 M) 0.4 L 2 L 10 L SYBR Green Realtime PCR Master Mix 1/2 ( ) 0.2 0.6 M (2)PCR 3 Tm 55 65 60 15 PCR 1 5 Channel F1 2 data collection [PCR ( )](3 60 72 ) 95 60 95 15 40 60 15 72 30 (data collection) (Melting Curve) - - 9 - -

1 RT-PCR (1) cdna DNA PCR RNA 1 RT-PCR 1 RT-PCR PCR ReverTra Ace (Code:TRT-101,102) (2) PCR RNase Inhibitor (Code:SIN-101,102) 0.5U/ L ReverTra Ace 0.01 0.3U/ L( ) ReverTra Ace ReverTra Ace RNase Inhibitor 5U/ L ReverTra Ace (RT ) RT RT DNA DNA RT(-) ( ) [RT ( )](ReverTra Ace 3U/ L ( 0.3U/ L)) RT (RNase Inhibitor 5U/ L) 32 L ReverTra Ace (100U/ L) 1 L 33 L [PCR ( )]( 0.4 M 50 L) 11 L SYBR Green Realtime PCR Master Mix 25 L 1 (10 M) 2 L 2 (10 M) 2 L RT (ReverTra Ace 3U/ L) 5 L 5 L 50 L SYBR Green Realtime PCR Master Mix 1/2 ( ) LightCycler TM 20 L LineGene 10 L 0.2 0.6 M - - 10 - -

(3)PCR PCR ( ) 42 20 5 ( PCR ) Tm 55 65 [ABI PRISM 7700( )] 42 20 95 5 95 15 40 60 15 72 45 (data collection) [Roche LightCycler TM ( )] 42 20 95 5 95 5 40 55 10 72 15 (data collection) [BioFlux LineGene( )] 42 20 95 5 95 15 40 60 15 72 30 (data collection) - - 11 - -

(1) ( ) SYBR Green I (2) PCR PCR GC cdna (1) ( ) (2) cdna - - 12 - -

PCR PCR GC (1) PCR ( ) (2) ( ) PCR 3 Code No. ReverTra Ace - - 100FSK-101 ReverTra Ace RNase Inhibitor Realtime PCR Master Mix 10,000U 1 50,000U 1 2,500U 1 2,500U 5 1mL 1 1mL 5 TRT-101 TRT-102 SIN-101 SIN-102 QPK-101T QPK-101 - - 13 - -

530-8230 2 8 TEL 06-6348-3786 FAX 06-6348-3833 E-mail : order_lifescience@bio.toyobo.co.jp 103-8530 17 9 TEL 03-3660-4819 FAX 03-3660-4951 E-mail : order_lifescience@bio.toyobo.co.jp TEL 06-6348-3888 FAX 06-6348-3833 9:00 12:00, 13:00 17:00 E-mail : techosk@bio.toyobo.co.jp URL http://www.toyobo.co.jp/seihin/xr/lifescience.html