The overall process is illustrated by the image to the right; a more detailed view is in the table below. In particular, it shows
Timings are very rough estimates. E.g. the first visit is indicated to take a week, whereas for a well-characterised system this can be completed in a day, if you start early. Experienced users also get through this far quicker than listed here.
Also, things may deviate significantly if the crystals demand it - and what they need is highest priority for these experiments.
|(Approximate timing)||User||Local Contact|
|Before scheduling first visit|| |
|Optimize crystallization for one of supported plate types (100% working: SwissCi/MRC 3-drop; in trial: MRC 2 wells and crystalQuickX)||Skype/phone call with user (and PBS): review science, experiment detail (crystalli, agree timelines, review experiment|
|Register with Diamond and the Research Complex. Access documents will be required and also completing video training on DLS website.||Create visit LBXYZ-1 so it starts 1 week before training week (the creation instructions are in confluence).|
|Confirm dates in UAS so accomodation can be booked.|| |
Make sure accomodation is booked 4 weeks in advance or more.
FOR EACH VISIT TO DIAMOND: Email the DLS user office, RCaH admin (Louise Hatter and Sonia Joiner) and DLS lab manager (Jams Sandy) for access to lab34a.
|Confirm consistent diffraction from 10-16 identically mounted apo crystals (either do experiment, or send historic images)||Review diffraction images and processing (email etc)|
|If bringing crystals: confirm that crystals travel|
If crystallising at Diamond: confirm that protocol is transferrable. Then to actually ship the samples:
|Receive protein/reagents/seeds, set up crystallization (Echo AND mosquito)|
|If new plate type: provide images of blank wells, and send empty plate to Local Contact|| |
|If own compounds: get into Echo-compatible plates.|
Schedule first visit:
|Week before visit|
Grow crystals (if bringing). Almost certainly better that they're fresh.
Register the known model PDB and the protein sequence in ISPyB.
|Grow crystals (if at Diamond)|
|Finalise travel arrangements|
|Day before visit|
Aim to arrive day before:
|Prepare lab: schedule conflicts; check loops, pucks, plates, fragments; tidy up|
(Or start Tuesday - but no beam till Weds)
Solvent test (usually DMSO)
get user's plates imaged (RCaH)
|Wednesday or Thursday|| |
|Thursday or Friday|| |
|Friday (wrap-up)|| |
1. (No harvesting: lab down day)
|1. () |
2. Review with user
3. Schedule next visit
4 Clean loops and lab
5. Fill summary spreadsheet
|Before Visit #2|
|Register visit (guest house!!)|
|Grow enough crystals (home or Diamond)|
Day before: Prepare lab
|Visit #2: Full run|
|Day 1||(lab) soak Nx100 fragments|
|Day 1 or 2||(lab) harvest Nx100 crystals||schedule beamline queue (EHCs?)|
crystals onto beamline queue (low)
|Present/remote||(computer) assess diffraction, analyse hits|
|Review results with Local Contact, PBS||Review results with user|
|(Further work as required)|
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