User:Mackenzie L. Cowell/IGEM 2007 part fab protocol

Overview

The 2007 DNA part kit will be comprised of around 1500 parts contained on 4 384-well plates. We need to fabricate 150 copies of the kit, producing a total of 600 384-well plates. The parts are contained on plasmids in cultures of E. coli frozen in glycerol stocks in 96-deep-well (96d) plates in -80 freezers at the registry. We are going to inoculate 96d plates from the freezer stocks and give them to Ed's lab for incubation and then miniprepping in 4 groups of 4, for a total of 16 96d plates. For every 4 that we get back we will use the EPM to compose the miniprepped DNA into 9 384d intermediate source plates (SP), of which we will give 3 back to Ed's lab to use with the PMP to transfer 3.5 uL of DNA into 150 384-well kit plates. This will happen 4 times, producing an uncollated set of the 600 384-well plates for the 150 07 DNA part kits.

Supply List

The 07 kits will be comprised of no more than 4 384-well plates (1536 parts), so to make 150 kits, we'll need:

• 32 96-deep-well square-bottom plates
  • 16 for Ed's miniprep, will be sources for the intermediate SPs
  • 16 for miniprepping for Robotic Assembly supplies
• 16 96-deep-well round-bottom plates (for 2 glycerol stocks of each new registry freezer plate)
• 20 384-deep-well plates
  • 16 for Antibiotic Testing
  • 4 for Robotic Assembly supplies
• 660 384-well plates, for kits (4 per kit)
  • 600 for kits (4 per kit)
  • 60 for intermediate Source Plates for kit fab with the PMP (the PMP 384 tip head can't reach the bottom of 384d plates)

• 672 foil covers
• 656 plastic covers (AB test plates don't need plastic covers & the plates come with them already)
• 15 rolls label tape
• 32 boxes of tips

(Items that are crossed out have been delivered to the Registry and are stored in 314)

Test Run: 10 April 07

Meet Sultrim around noon with 6 384 plates filled with 100 uL dyed dH20 per well (230.4 mL ) and 20 empty 384 well plates. Try filling the empty plates with 3.0 uL from the SPs.