PCR(KODplus)

Preparation

• Components Volume Final Concentration
• 10× PCR Buffer for KOD -Plus- 5 μl 1×
• 2mM dNTPs 5 μl 0.2mM each
• 25mM MgSO4 2 μl 1.0mM
• Primer (10μM each) 1.5 μl 0.3μM each
• Template DNA ≧ 1 μl
• Genomic DNA 10~200 ng/50μl
• （Plasmid DNA 1~50 ng/50μl）
• KOD -Plus- (1U/μl) 1 μl 1 U/50μl
• MiliQ to 50 μl

PCR program

• Pre Denature : 94 degrees, 2min.
• Denature : 94 degrees, 15sec.
• Annealing : (Tm-5) degrees, 30sec.
• Extension : 68 degrees, 1min. /kb
• Set extension time for 1 min./kb