KOD －Plus－ Ver．2
Contents
1. gt TOYOBO TEL 06 6348 3888 FAX 06 6348 3833 9 00 lt 12 00 13 00 lt 17 00 E mail tech_osaka toyobo jp URL http www toyobo co jp bio A3641K HPCR 3 KOD Plus Ver2 HeLa total RNA DNA Polymerase EcDNA 6 8kb 4 PCR 1 PCR Components Volume FinalConcentration 10x Buffer for KOD Plus Ver 2 5 gl 1x 2mM dNTPs 5 gl 0 2mM each 25mM MgSO4 3 al 1 5mM 10 uM each 1 5 0 3Meach Genomic DNA 10 200 ng50 DNA iul Plasmid DNA 1 lt 50 ng50 cDNA 5 KOD P2. RT PCR Taq DNA Polymerase 80 KOD Plus PCR KOD KOD Plus TA Arget Clone Plus KOD Plus Ver2 KOD Plus Buffer 10xBuffer for KOD Plus KOD Plus KOD Plus Ver2 Buffer Code No KOD 3B KOD Plus Ver2 1 KOD 211 KOD 211X5 200Ux1 KOD Plus 1 0 U l 200lx1 10x Buffer for KOD Plus Ver 2 1mlx1 25mM MgSO 1mlx1 NN 2mM dNTPs 1mlx1 2
3. PCR 12 PCR blunt end PCR blunt end KOD KOD Plus TA TArget Clone Plus Code No TAK 201 TA MgSO 1 5mM 2 0mM Tm 10 C 2 lt 5 DNA EE 1U 1 5 2 0U DNA DNA RNA GC rich MgSO 1 5mM 1 0mM RNA cDNA
4. RNA 68 C a 2 PCR SO 1 5mM 1 0mM 2 5 DNA MgSO 1 5mM 1 0mM 1U 0 5 0 8U TA TArget Clone Plus gt TOYOBO TEL 06 6348 3888 FAX 06 6348 3833 9 00 lt 12 00 13 00 lt 17 00 E mail tech_osaka toyobo jp URL http www toyobo co jp bio HPCR 7 1 CDNA KOD Plus PCR
5. HeLa total RNA cDNA PCR KOD Plus Ver2 human polymerase cDNA 6 8kbp 4 KOD Plus Ver2 KOD Plus PCR M 1 2 3 4 1 2 3 4 1 2 3 4 1 human transferrin receptor cDNA 3 5kb 2 human tuberous scleosis CDNA 5 3kb 3 human adaptn cDNA 5 7kb 4 human polymerase cDNA6 8kb M 1kb ladder VOL2 gt TOYOBO i 9 00 12 00 13 00 17 00 E mail tech_osaka toyobo jp URL http www toyobo co jp bio 8 Code No 2 000 Ux1 TRT101T lt M MLV gt 10 000 Ux1 TRT101 ReverTra Ace 100 U l 10 000 Ux1 x5 TRT 101
6. PCR 50 RT RNA 150ng Total RNA1 gg 20 RT PCR 3 DNA RNA RNA DNA DNA RNase 8 GC rich DMSO 2 gt 59 9 DNA 94C 15sec GC rich 98 C 10sec 10 PCR DNA DNA 50 200ng 30 lt 40 11 Colony Direct PCR PCR
7. Mixture 2mM Code No NTP 201 4 GC 22 34mer Tm gt 60C 5 PCR Tm 65C 6 DNA PCR DNA RNA PCR gt TOYOBO TEL 06 6348 3888 FAX 06 6348 3833 9 00 lt 12 00 13 00 lt 17 00 E mail tech_osaka toyobo jp URL http www toyobo co jp bio 7 DNA RT RNA PCR
8. 14 04 HPCR Code No KOD 211 Code No KOD 211X5 2 0 PCR KOD Plus Ver 2 Thermococcus kodakaraensis KOD1 KOD DNA Polymerase PCR Polymerase 5 Exonuclease ProofHreading Taq DNA Polymerase 80 KOD Plus PCR Buffer PCR KOD Plus Polymerase 5 Exonuclease 2 Hot start PCR PCR RT KOD Plus
9. 3888 FAX 06 6348 3833 9 00 lt 12 00 13 00 lt 17 00 E mail tech_osaka toyobo jp URL http www toyobo co jp bio 530 8230 2 8 TEL 06 6348 3786 FAX 06 6348 3833 E mail order lifescience toyobo jp 104 8345 17 10 TEL 03 6887 8819 FAX 03 6887 8951 E mail order lifescience toyobo jp gt TOYOBO TEL 06 6348 3888 FAX 06 6348 3833 9 00 lt 12 00 13 00 lt 17 00 E mail tech_osaka toyobo jp URL http www toyobo co jp bio
10. X5 10 000 Ux1 x10 TRT 101X10 lt PCR gt 250Ux1 KOD 101 KOD DNA Polymerase 250Ux1 x5 KOD 101X5 200Ux1 KOD 201 a 0 200 Ux1 x5 KOD 201X5 200 Ux1 x10 KOD 201X10 lt KOD KOD Plus TA gt TArget Clone Plus ME JAN dNTPs Mixture 2mM 1 ml NTP 201 KOD Plus 25mM MgSO 1ml KOD 2S KOD Plus Ver2 Buffer 1 ml KOD 3B 9 1 Takagi M Nishioka M Kakihara H Kitabayashi M Inoue H Kawakami B Oka M and Imanaka T App Environ Microbiol 63 4504 4510 1997 2 Hashimoto H Matsumoto T Nishioka M Yuasa T Takeuchi S Inoue T Fujiwara S Takagi M Imanaka T and Kai Y J Biochem Tokyo 125 983 986 1999 3 Mizuguchi H Nakatsuji M Fujiwara S Takagi M and Imanaka T J Biochem Tokyo 126 762 768 1999 4 Nishioka M Mizuguchi H Fujiwara S Komatsubara S Kitabayashi M Uemura H Takagi M and Imanaka T J Biotechnol 88 141 149 2001 5 Hashimoto H Nishioka M Fujiwara S Takagi M Imanaka T Inoue T and Kai Y J Mol Biol 306 469 77 2001 6 Imanaka T and Takagi M J Chin Inst Chem Engrs 32 277 288 2001 gt TOYOBO TEL 06 6348
11. lus 1U 1 gl 1U 50l Autoclaved distilled water to 50 gl 2 PCR 3 Tm 65C PCR 2 3 2 Predenature 94 C 2min Predenature 94C 2min Denature 98 C 10sec Denature 98C 10sec 25 40 Annealing Tm 5 C 30sec NE Extension 68C 1mn kb cycles Extension 68 C 1min kb Extension 1kp 1min 5 1 thin wal PCR total 50 ux I 2 3 dNTPs PdNTPs
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