title: USDA Bradyrhizobium japonicum 8.5K (BJ_USDA110_Oligo_Array)
status: Public on Jul 15 2007
contact.city: Arlington
contact.first: Woo-Suk
contact.last: Chang
contact.email: wschang@uta.edu
contact.state: TX
contact.address: 501 S. Nedderman Dr. , Life Science Building 216
contact.department: Biology
contact.country: USA
contact.institute: University of Texas at Arlington
contact.phone: 1-817-272-3280
organism: Bradyrhizobium japonicum
manufacture_protocol: 1. Unique 70-mer oligonucleotides were designed and synthesized for each of the predicted ORFs by Operon Biotechnologies, Inc. (Huntsville, AL). 2. Arrayer pre-printing setup - Remove any large objectives that would obstruct the arrayer's path from the deck. - Clean the pins for 15 min with warm (40 celsius) water followed by washing with 0.5% SSC. - Dry the pins. 3. Microarray printing: Oligonucleotides were printed onto Corning® epoxy coated slides (Corning Inc., Lowell, MA) at the Washington University Genome Sequencing Center (St. Louis, MO). - Arraymaker 2 software was used. - Set up: 2X wash to 4500 msec. - Remove the orange rubber strips and wipe off the columns needed to print on the deck with a 95% isopropanol swipe and allow it to dry. - Place orange rubber strip, located next to the deck, over all other unused columns on the deck. - Place the slides in their appropriate slide locations. - Place the first 384 well plate into the centrifuge and spin it at 1,500 rpm for 1 minutes at room temperture. - Place the spun down plate in the holder with appropriate position. Make sure the plate is sitting flat. - Make sure all the slides are moved toward the left of the column and to the top of their columns for uniformity of printing. - Plug in the pump for the deck. - Click on the "START" button. 4. Post processing slides Ultra GAPS - Put the printed slides into a slide box and store them in a dessicant cabinet for 48 hrs. - Add 1.5 L of MilliQ water to a pyrex container and heat the water until steaming. - Take out a slide and hold it over the steaming water, element slide down, and allow the slide to cover with steam for 8-10 seconds. - Immediately place the slide, element up, on the heat block for 8-10 seconds to allow the slide to snap dry. - After rehydration, place them in the UV stratalinker. - Turn on the machine and press in "3000". Make sure 3000 appears on the digital screen (300mJ). - Press energy button and then press the start button. - After crosslinkin, scribe each slide with a diamond scribe and place the slides in a slide box. - Store them in a dessicant cabinet.