title: Biomphalaria glabrata 70-mer oligo array version 1
status: Public on Jan 01 2010
contact.city: Albuquerque
contact.first: Patrick
contact.middle: C
contact.last: Hanington
contact.email: pch1@unm.edu
contact.state: NM
contact.address: 1 University of New Mexico
contact.department: Department of Biology, University of New Mexico
contact.country: USA
contact.institute: Center for Evolutionary and Theoretical Immunology
organism: Biomphalaria glabrata
manufacture_protocol: The 1152 oligonucleotides supplied by IDT, originally provided at a 6nM concentration were resuspended in deionized water to a concentration of 100μM. They were further diluted into 50% water and 50% ESB (Epoxide Spotting Buffer, Integrated DNA Technologies, Coralville, IA) to a final concentration of 40μM for spotting on the microarray. Twenty microliters of each oligonucleotide was transferred to 384 well microarray plates (Genetix, Queensway, New Milton, Hampshire, UK). A total of 300 Epoxide-coated glass slides (Corning Life Sciences, Lowell, MA) were spotted using the QArray Max (Genetix, Queensway, New Milton, Hampshire, UK) and dried for one hour at 80% humidity and then overnight at 60% humidity. A 24 pin microarraying head was used to print the slides. The plate type selected was for a 10μl calibration even though there was 20μl of sample per slide. Eight landing lights (consisting of 5μM Cy5 labeled oligo, 5μM Cy3 labeled oligo, 40μM of unlabeled oligo (GCF3), and ESB) were printed on every subarray for orientation. The printing conditions consisted of 630 stamps/ink and 1 stamp/spot. The stamp time for each spot was 20ms (milliseconds) and the total inking time was 9999ms. A total of 2 touch-offs per inking were used and lasted 4000ms. The sterilization cycle used included 10000ms wash time and 35000ms dry time. All samples were printed first and the markers printed last.Three replicates of each oligo were printed in each field, resulting in 6 replicate spots per slide. Replicates were spotted so that they were not adjacent to each other. A total of 48 subarrays with dimensions 12 x 13 spots were printed on each slide resulting in a 7,488 feature array. All slides were stored in a dessicated mircro array cabinet until use.