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From 2012.igem.org

Contents 1 Methods 2 Protocol 2.1 Medium & Buffer recipes 2.2 CCMV Coat Protein VLP formation 2.3 Hepatitis B Coat Protein VLP formation 2.4 TuYV Coat Protein VLP formation 2.5 PLRV Coat Protein VLP formation 2.6 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.

• The production of our VLPs monomers is done in E.coli, while the formation of our VLPs is done in vitro. We used multiple techniques to make this happen. Click here to read more
• After the production of our VLPs we have our formed VLPs, but this product is not pure enough. So we developed a two step method to purify the product. Click here to read more

Protocol

Medium & Buffer recipes

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

CCMV Coat Protein VLP formation

• Growing culture
• Dialysis of the VLPs
• Purifing the VLPs

Hepatitis B Coat Protein VLP formation

• Growing culture
• Dialysis of the VLPs
• Purifing the VLPs

TuYV Coat Protein VLP formation

PLRV Coat Protein VLP formation

• RNA isolation from potato leaf material

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