title: VIB Bacillus thuringiensis 1K VIB_Bt_1K_1
status: Public on Sep 23 2008
contact.city: Leuven
contact.first: Rekin's
contact.last: Janky
contact.email: Nucleomics.Bioinformatics@vib.be
contact.address: Herestraat 49 Box 816
contact.department: Nucleomics Core
contact.country: Belgium
contact.institute: VIB
organism: Bacillus thuringiensis
description: The array design contains 672 features printed in a layout of 4 meta rows and 4 meta columns; consisting out of 7 rows and 6 columns per block. The array measures the expression of 150 unique sequences, spotted with four replicates. A set of 72 spots remained empty.
  The platform data table reflects a condensed representation of the array. The complete array layout file representing the replicated features as well is linked as a supplementary file.
manufacture_protocol: Equipment:
  1. Microgrid II (Genomic Solutions, Huntingdon, United Kingdom).
  2. 384 microtiter plates (Genetix)
  3. 16 MicroSpot quill pins - 10K (cat# BR001489, Genomics Solutions, UK)
  4. Codelink slides (cat#: 30001100, GE Healthcare).
  Printing 2 arrays of 4x4 blocks on 1 microscope slide:
  1.  All oligos were obtained from Sigma Genosys and dissolved in H2O to a concentration of 100µM. Subsequently, oligos were diluted with a 156.25mM Na-Phosphate buffer plus 0.00015625% sarcosyl to a final concentration of 20 µM.  Re-array the oligos in 384 microtiter plates.
  2. Set spotter settings for 2 arrays of 4x4 blocks printed with 16 pins: column pitch = 190 um, row pitch = 190 um; Humidity 55-60%; 123 spots per source visit.
  3. Make sure room temperature is between 20-24 degrees Celsius.
  4. Place microtiter plates in the plate hotel with A1 in front left corner.
  5. Place 25 slides on the slide-deck.
  6. After spotting, incubate the slides in humid chamber with saturated NaCl for min 24 hours.
  7.  Slides were treated with 1% NH4OH solution on a shaker for 5 min, followed by 0.1% SDS for 5 min. The slides were rinsed in MQ-water, placed in 95°C water bath for 2 min, and then at -20°C for 1 min. The slides were rinsed again twice in MQ-water and finally dried by centrifugation (1min at 1000rpm). These slides were kept in the incubator at 24°C.