title: Earthworm 4K cDNA array version 1.0
status: Public on Oct 15 2007
contact.city: Vicksburg
contact.first: Ping
contact.last: Gong
contact.email: ping.gong@us.army.mil
contact.state: MS
contact.address: 3909 Halls Ferry Road
contact.department: Environmental Laboratory
contact.country: USA
contact.institute: US Army ERDC
contact.phone: (601) 634-3521
organism: Eisenia fetida
manufacture_protocol: 1. A total of 4032 purified cDNA clones were loaded on 384-well plates and completely dried down in a Vacufuge Concentrator 5301 (Eppendorf, Westbury, NY).
  2. The dried cDNA was re-suspended in 15 ?̬ of 1??printing buffer (ArrayIt, Sunnyvale, CA).
  3. Each clone was spotted twice (i.e., in two super grids) on Ultra GAPS amino silane coated glass slides (Corning, Acton, MA) using 16 pins on a VersArray ChipWriter (Bio-Rad, Hercules, CA).
  4. Five alien cDNAs, i.e., PCR product 1 to 5 selected from SpotReport Alien cDNA Array Validation System (Stratagene, La Jolla, CA) prepared at 15, 30, 60, 125 and 250 ng/?̬, were also spotted twice along with printing buffer and water as control spots.
  5. A total of 4032 purified cDNA clones were loaded on 384-well plates and completely dried down in a Vacufuge Concentrator 5301 (Eppendorf, Westbury, NY).
  6. The total number of spots on each array was 8704 including 60 alien cDNA spots, 84 water spots, 256 blank spots and 240 printing buffer spots.
  7. After printing, arrays were incubated in a dessicator 2-3 days and were then snap-dried on a hot plate after being rehydrated over a boiling water bath.
  8. The arrays were further immobilized using a UV Cross-linker (Stratagene) by applying 300 mJ per 10 arrays.