Where possible, I will upload relevant protocols here.
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.
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.
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.
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.
Protocol for golden gate assembly used in the Mackinder lab. Kindly typed up by visiting rotation student Mel Ludwig.
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.
Protocol for fluorescent labelling of purified proteins using Atto NHS ester dyes. This protocol has been used successfully for a range of proteins at concentrations down to ~0.4 mg mL-1. The addition of bicarbonate immediately prior to labelling improves reaction efficiency by increasing the pH into the optimal range for NHS ester chemistry.