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Team:UT-Tokyo/PCR（KODplus）
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| - | Pre Denature : 94 degrees, 2min. | + | *Components Volume Final Concentration |
| - | Denature : 94 degrees, 15sec. | + | *10× PCR Buffer for KOD -Plus- 5 μl 1× |
| - | Annealing : (Tm-5) degrees, 30sec. | + | *2mM dNTPs 5 μl 0.2mM each |
| - | Extension : 68 degrees, 1min. /kb | + | *25mM MgSO4 2 μl 1.0mM |
| - | Set extension time for 1 min./kb | + | *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 | ||
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| + | *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 | ||
{{UT-Tokyo_Foot}} | {{UT-Tokyo_Foot}} | ||
Revision as of 08:04, 23 September 2010
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
- 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
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