From 2012.igem.org

Revision as of 11:12, 26 October 2012

• Home
• Team
  • The Team
  • Team Registration
• Project
  • 1. Introduction
  • 2. PnA System
  • 3. Virus-Like Particles
    • CCMV
    • HepB
    • Polerovirus Natural BioBrick
  • 4. Outside Modification
  • 5. Inside Modification
  • 6. Detection of VLPs
  • 7. Applications
  • 8. The Constructor
  • 9. Final Overview
• Human Practices
  • Human Practices Home
  • 1. Mini Symposium
  • 2. Visiting Secondary Schools
  • 3. Discovery Festival
    • Communication Science
  • 4. Stakeholders
  • 5. Munich CAS Conference
• Safety
  • Introduction
  • 1. General safety
  • 2. Virus-related safety
  • 3. Regulations
  • 4. Safety suggestions
  • 5. Safety of applications
• Modeling
  • Human Body Model
  • Tertiary folding prediction
  • VLP assembly model
• Attributions
• Parts
• Notebook
  • Journal
  • Protocols

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Last year's project: Synchronized Oscillatory System

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Mutagenesis

QuikChange® Lightning Site-Directed Mutagenesis Kit

Prepare the sample reaction(s) as indicated below

1. 5 μl of 10× reaction buffer
2. X μl (10–100 ng) of dsDNA template
3. X μl (125 ng) of oligonucleotide primer #1
4. X μl (125 ng) of oligonucleotide primer #2
5. 1 μl of dNTP mix
6. 1.5 μl of QuikSolution reagent
7. ddH2O to a final volume of 50 μl
8. Then add: 1 μl of QuikChange® Lightning Enzyme

Cycling Parameters for the QuikChange Lightning Site-Directed Mutagenesis Method

* For example, a 5-kb plasmid requires 2.5 minutes per cycle at 68°C.