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From 2012.igem.org

Contents 1 Methods 2 Protocols 2.1 Medium & Buffer recipes 2.2 CCMV Coat Protein VLP formation 2.3 Hepatitis B Coat Protein VLP formation 2.4 Polerovirus Coat Protein VLP formation 2.5 General Protocol

Methods

The use of Virus-Like-Particles as medicine carrier is new for iGEM. This means the whole production, purification and detection of Virus-Like-Particles is also new in iGEM. In this section we will explain how the different methods work and how it all fits together.

• Production
• Purification

Protocols

Medium & Buffer recipes

• LB medium
• SOB medium
• SOC medium
• Disassembly buffer
• Dialysis buffer 10x
• Reassembly buffer
• Virus buffer
• Formation Buffer HepBcAg
• Washing Buffer HepBcAg
• Formation Buffer Polero
• Towbin's electrotransfer buffer 1x
• Towbin's electrotransfer buffer 10x

CCMV Coat Protein VLP formation

• Growing culture
• Dialysis of the VLPs
• Purifying the VLPs

Hepatitis B Coat Protein VLP formation

• Growing culture
• Dialysis of the VLPs
• Purifying the VLPs

Polerovirus Coat Protein VLP formation

• RNA isolation from potato leaf material
• Growing culture
• Dialysis of the VLPs
• Purifying the VLPs

General Protocol

• Preparation of electrocompetent cells of E. coli
• SDS-Polyacrylamide and native gel electrophoresis
• Dynamic Light Scattering user manual
• French press user manual
• FPLC user manual
• Mutagenesis