title: Maize long oligonucleotide arrays 57K, version 1.13 Array A
status: Public on Jan 01 2010
contact.city: Irapuato
contact.first: June
contact.last: Simpson-Williamson
contact.email: jsimpson@ira.cinvestav.mx
contact.state: Guanajuato
contact.address: Km 9.6 Libramiento Norte Carretera Irapuato-Leon
contact.department: Plant Genetic Engineering
contact.country: Mexico
contact.institute: Centro de Investigacion y de Estudios Avanzados del Instituto Politecnico Nacional
contact.phone: (0052) 462 6239600
organism: Zea mays
description: similar to Platform GPL5439
manufacture_protocol: REHYDRATION AND PRINTING OF THE OLIGONUCLEOTIDE MICROARRAYS  1.Add 15 uL of 3XSSC to each well of the 300 pM oligonucleotide set to make a 20 μM oligonucleotide solution.  2.Transfer onto an orbital shaker for 1 h at RT at 100 rpm. The oligonucleotides are now ready for printing.  3.The Arabidopsis oligonucleotide microarrays are printed on aminosilane-coated “Superamine” slides (Telechem) or “Powermatrix” slides (Full Moon Biosystems), using Telechem Type SMP3 printing pins.  4.Printing conditions:  Single array element printed per oligonucleotide, 48 pins in the print head, 31% relative humidity, redipping after every 50 slides. The print head Z- velocity is set to a maximum of 10 mm/sec.  5.Bake slides for 2h at 80oC. Store dry and in darkness at RT.  The microarrays are sent out to recipient scientists at this point. Prior to hybridization, these next steps should be followed:  IMMOBILIZATION OF THE OLIGONUCLEOTIDE ARRAY ELEMENTS    1.Rehydrate the microarray slide over 50oC water for 5-10 seconds; snap-dry on a 65oC heating block for 5 seconds. Repeat this step 4-6 times, examining the slides under a dissection microscope (or using red reflectance imaging) to verify complete re-hydration of all the spots. If re-hydration is not complete, repeat the process several more times.  2.Crosslink the oligonucleotides to the slide surface by exposing the DNA-side of the microarray to UV irradiation. We employ a Stratalinker set at an energy level of 180 mJ.    Re-hydration and UV cross linking can be done well in advance before microarray hybridization, and the slides can be stored at room temperature for several months. However we do not recommend storage of washed microarray slides.  3.Wash the slides in 1% SDS dissolved in sterile ddH2O for 5 min at RT.  4.Rinse the slides by plunging 10 times into sterile ddH2O.  5.Rinse in 100% ETOH for 30 seconds to 5 min, with gentle shaking.  6.Spin dry the slides in a centrifuge at 1000 rpm for 2 min.  7.Store the slides in a light-proof box under cool dry conditions.