Protocols

Where possible, I will upload relevant protocols here.

• Chloroplast transformation by biolistic bombardment

  Standard protocol for the transformation of the Chlamydomonas reinhardtii chloroplast using biolistic particle bombardment. Adapted from a protocol shared by the lab of Alison Smith (University of Cambridge). Used for the transformation of Chlamydomonas in a recent Mackinder lab publication, supported by work from collaborators.

  
  
• Pyrenoid enrichment protocol

  Protocol for the enrichment of pyrenoids from green algae. Used for the isolation of pyrenoid enriched fractions from Chlorella sorokiniana in a recent Mackinder lab contribution.

  
  
• Rubisco purification protocol (sucrose gradient)

  Protocol for the high quality purification of Rubisco, applied to green algae and plants, yielding active, intact Rubisco holoenzyme complexes. This protocol was used for purification of all Rubiscos in a recent Mackinder lab publication.

  
  
• Linker purification protocol

  Protocol for the soluble purification of linker proteins from algal pyrenoids. Used for the purification of Chlamydomonas (EPYC1) and Chlorella (CsLinker) linker proteins, as well as a host of other soluble algal pyrenoid proteins.

  
  
• Golden gate assembly protocol

  Protocol for golden gate assembly used in the Mackinder lab. Kindly typed up by visiting rotation student Mel Ludwig.

  
  
• Rubisco purification protocol (15Q)

  Protocol for the high quality purification of Rubisco, applied to green algae, seaweed and plants, yielding active, intact Rubisco holoenzyme complexes. This protocol was developed from an original protocol that included sucrose gradient centrifugation, which was limiting for the throughput of the purification. In this protocol, a 15Q ion exchange column is used. Lower quality ion exchange resins (e.g. HiTrap Q XL) are not appropriate for this.