Transcription of KOD －Plus－ Neo - TOYOBO
1 (KOD-401) TEL 06-6348-3888 FAX 06-6348-3833 9:00~12:00, 13:00~17:00 ( ) E-mail: [URL] Code No. KOD-401 Code No. KOD-401X5 Code No. KOD-401X10 C PCR KOD - plus - Neo Thermococcus kodakarensis KOD1 KOD DNA Polymerase1,2) PCR KOD DNA Polymerase 3 5 Exonuclease Proofreading PCR KOD DNA Polymerase KOD - plus - Code No. KOD-201 KOD - plus - Ver. Code No. KOD-211 Taq 80 1 kb 30 kb PCR Polymerase 3 5 Exonuclease 2 Hot start PCR KOD - plus - Neo * Taq 80 *KOD DNA polymerase PCR 20~30 < > < > 20mer Tm * 63 C 2 22-05 * Tm Nearest Neighbor method Tm 50 mM Na+ M
2 (KOD-401) TEL 06-6348-3888 FAX 06-6348-3833 9:00~12:00, 13:00~17:00 ( ) E-mail: [URL] 1 kb 30 kb DNA kb Mg 2 mM DNA kb KOD- plus - Neo 3 KOD-401 (200 U 1 ) KOD-401X5 KOD-401X10 KOD - plus - Neo ( U/ L) 200 L 1 (KOD-401) 5 (KOD-401) 10 10 PCR Buffer for KOD - plus - Neo 1 mL 1 25 mM MgSO4 1 mL 1 2 mM dNTPs 1 mL 1 KOD - plus - Neo DNA -globin kb (KOD-401) TEL 06-6348-3888 FAX 06-6348-3833 9:00~12:00, 13:00~17:00 ( ) E-mail.
3 [URL] PCR 22~35mer Tm * 63 C GC 45~60% GC GC 3 5 60~70%, 3 40~50% GC 3 G C 3 GC 25~35mer Tm 65 C 25mer Tm 65 C * Tm Nearest Neighbor method Tm 50 mM Na+ M Nearest Neighbor method Tm ( ) PCR Components Volume Final Concentration Autoclaved, distilled water (33 - X) L 10 PCR Buffer for KOD - plus - Neo 5 L 1 2 mM dNTPs 5 L mM each 25 mM MgSO4 3 L mM (10 M each) L M each X L Genomic DNA ~200 ng/50 L Plasmid DNA ~50 ng/50 L cDNA ~200 ng/50 L KOD - plus - Neo (1 U/ L)
4 1 L 1 U/50 L Total 50 L PCR Buffer 1 Mg mM 10 kb M 10 kb M 2 RNA PCR PCR 50 L RNA 50 ng Total RNA1 g 20 L 1 L 5. 1 (KOD-401) TEL 06-6348-3888 FAX 06-6348-3833 9:00~12:00, 13:00~17:00 ( ) E-mail: [URL] PCR [ ] KOD - plus - Neo 2 Tm 63 C Tm 68 C 2 A.
5 2 Tm 63 C 2 2 Predenature : 94 C, 2 min. Denature : 98 C, 10 sec. Extension : 68 C, 30 kb 30~45 Extension 1 kb 30 sec. 10 kb 1 kb Mg 2 mM DNA Denature 94 C, 15 sec. GC rich 98 C, 10 sec. DMSO 3 25~35mer Tm 68 C Tm 68 C 2 DMSO 2% 25 45 cycles (KOD-401) TEL 06-6348-3888 FAX 06-6348-3833 9:00~12:00, 13:00~17:00 ( ) E-mail: [URL] B. Tm 63 C 2 3 3 Predenature : 94 C, 2 min.
6 Denature : 98 C, 10 sec. Annealing : (Tm) C, 30 sec. Extension : 68 C, 30 kb Tm Predenature : 94 C, 2 min. Denature : 98 C, 10 sec. Extension : 74 C, 30 kb Denature : 98 C, 10 sec. Extension : 72 C, 30 kb Denature : 98 C, 10 sec. Extension : 70 C, 30 kb Denature : 98 C, 10 sec. Extension : 68 C, 30 kb Extension : 68 C, 7 min. 3 10 kb 1 kb Mg 2 mM DNA Denature 94 C, 15 sec. GC rich 98 C, 10 sec. 15 30 cycles 5 cycles 5 cycles 5 cycles 25 45 cycles (KOD-401) TEL 06-6348-3888 FAX 06-6348-3833 9:00~12:00, 13:00~17:00 ( ) E-mail.
7 [URL] PCR PCR 50 L Genomic DNA DNA 5~200 ng 50 ng DNA ~100 ng 10 ng Plasmid DNA 10 pg~50 ng 1 ng cDNA ~200 ng (RNA ) 50 ng (RNA ) DNA 10 pg~10 ng 1 ng PCR RNA PCR RNA PCR PCR 50 L RNA 50 ng Total RNA1 g 20 L 1 L GC rich DMSO 2~5% DMSO DMSO 3 25~35mer Tm 68 C Tm 68 C 2 DMSO 2% thin-wall PCR total 50 L dNTPs dNTPs Mixture 2 mM Code No.
8 NTP-201 (KOD-401) TEL 06-6348-3888 FAX 06-6348-3833 9:00~12:00, 13:00~17:00 ( ) E-mail: [URL] PCR PCR blunt end PCR PCR blunt end TA KOD DNA Polymerase TA TArget CloneTM - plus - Code No. TAK-201 PCR TA TArget CloneTM - plus - 10 A-attachment Mix Code No. TAK-301 PCR 3 A T PCR TA Ligation high Ver. Code No. LGK-201 PCR DNA Polymerase 3 5 Exonuclease / DNA MagExtractor TM -PCR & Gel Clean up- Code No.
9 NPK-601 (KOD-401) TEL 06-6348-3888 FAX 06-6348-3833 9:00~12:00, 13:00~17:00 ( ) E-mail: [URL] DNA -globin PCR TArget CloneTM - plus - Code No. TAK-201 TA 96 KOD - plus - Neo KOD - plus - KOD - plus - Ver. Taq DNA Polymerase 80 PCR KOD - plus - Neo PCR DNA -globin PCR 30 KOD - plus - Neo kb kb PCR HeLa Total RNA cDNA Total RNA ng 4 PCR 40 kod plus - Neo 4 (KOD-401) TEL 06-6348-3888 FAX 06-6348-3833 9:00~12:00, 13:00~17:00 ( ) E-mail.
10 [URL] DNA 50 ng -globin kb KOD - plus - Neo 30 kb 1 Total RNA ORF HeLa Total RNA cDNA Total RNA 50 ng ORF open reading frame PCR 30 KOD - plus - Neo (KOD-401) TEL 06-6348-3888 FAX 06-6348-3833 9:00~12:00, 13:00~17:00 ( ) E-mail: [URL] 2 DNA 200 ng -globin kb tPA Human tissue-type plasminogen activator kb 10 kb M (1) Takagi, M.
