title: Keasling Lab_In-house spotted 70-mer oligo_E. coli array
status: Public on Oct 01 2009
contact.city: Emeryville
contact.first: Robert
contact.last: Dahl
contact.email: robdahl@berkeley.edu
contact.state: CA
contact.address: 5885 Hollis St, JBEI
contact.department: Chemical Engineering
contact.country: USA
contact.institute: University of California-Berkeley
organism: Escherichia coli
manufacture_protocol: 1.  70-mer oligos (Operon - v.1.0.2) representing each ORF of E. coli were arrayed in 384 well plates, total of 600 pmol in each well.
  2.  Oligos were suspended in water to 40 uM, and 5 uL were transferred to a new 384 well plate.
  3.  The print plates were dried down in a speedvac and resuspended in 5 uL 3x SSC buffer, the day before printing.
  4.  Plates were sealed until just before use to prevent evaporation.
  5.  Super Amine 2 substrates (Telechem) were arrayed on a robotic spotter (Gene Machines).
  6.  The print run was designed to print each gene on a slide 3 times.
  7.  After printing, the print plates were dried in a speed-vac and stored at -80 C for storage.  For future printing, the plates were re-suspended in water.
  8.  Printed slides were rehydrated over water and UV cross-linked with 150 mJ/cm2.