ISISBio:Internal/Group meetings/13-June-08

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Safety Issues:

  • Autoclave is currently being quoted for plumbing in which seems to be getting out of hand from a cost perspective.
  • Up to two days disruption to fit tank in ceiling
  • Query over status of bunsen burner, some suggestion that it needs to be protected and clamped down. This isn't really practical.
  • Use of chloroform to clean troughs
    • Action LW to ask for risk assessment for trough use

Current Projects Status:

  • Preparation of protein for QENS
    • MT - lots of GFP available, needs to be purified by gel filtration in advance of beamtime
    • Need to not spend too much more time on this
      • Action CN to confirm with MT when the beamtime is, prefer September
  • Preparation of mellitin for VGS
    • Currently checking to see whether initial plasmid has been made
    • Several more mol biol steps to go
    • Action LW to follow through with next steps
  • OmpF for SAH
    • LC has purified significant quantities of OmpF, total of about 6 mg
    • Played around with making vesicles. PE doesn't make very good vesicles so need to move 25:25:50 PE:PG:PC vesicles
    • Going to Oak Ridge for two weeks from 29 June -9 July to deuterate OmpF for SANS experiments
    • May need to 5mL DEAE column to Oak ridge for final wash stage
    • External reflection IR and DLS organised to do at Reading
  • Ion channels
    • Currently trying to purify MLok1 in large quantities for ILL on 19 July
    • Recent issues with funny bands on gels which seemed to appear in last two weeks. Checking batches against each other.
    • Some sample to take but possibly not as much as we want. Would like 3mg/mL for 5/6 contrasts.
    • Will also take some mthK to ILL to see whether can see change on addition of Ca2+
  • Fluorescent labelling
    • Think Michael had made pcr product,now need to purify and ligate to make plasmid

Next week

  • Luke using big FPLC all week
  • CN/Maria to use baby Akta end of week (Wed-Fri) probably
  • MS preparing and transforming Mlok1-LPETGG plasmid and recloning sortase
  • VGS probably continuing with mol biol

Week after

  • Dave Barlow/Damian experiment to express and purify GlpF/DCytB
    • Action CN to contact DB and suggest moving schedule up to avoid Thursday


  • Need emergency protocol for MCR
  • Booking system for lab and equipment