Team:UT-Tokyo/PCR（KODplus）

From 2010.igem.org

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(New page: __NOTOC__{{UT-Tokyo_CSS3}}{{UT-Tokyo_Head}} <h1>PCR(KODplus)</h1> Components Volume Final Concentration 10× PCR Buffer for KOD -Plus- 5 μl 1× 2mM dNTPs 5 μl 0.2mM each 25mM MgSO4 2 μ...)
 
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<h1>PCR(KODplus)</h1>
<h1>PCR(KODplus)</h1>
-
Components Volume Final Concentration
+
<h2>Preparation</h2>
-
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.  
+
*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
 +
 
 +
<h2>PCR program</h2>
 +
*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}}

Latest revision as of 08:05, 23 September 2010

UT-Tokyo

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

Retrieved from "http://2010.igem.org/Team:UT-Tokyo/PCR%EF%BC%88KODplus%EF%BC%89"