Printing by the National Cancer Institute/Advanced Technology Center Microarray Facility Surface type: poly-L lysine coated glass slides Attachment: UV cross-linked at 600 mJ Array description: The long oligo (60-65 nt) custom arrays were manufactured at the Advanced Technology Center Microarray Facility (NIH/NCI/CCR/ATC). Microarrays were printed with a GeneMachines robotic arrayer from Genomic Solutions (http://www.genomicsolutions.com) that was fitted with a printhead containing four (4) SMP3 printing pins from Telechem International (http://www.arrayit.com). In addition to the KSHV probes, the array included human housekeeping genes for normalization purposes. The human housekeeping 65 nt oligomer set (Cat # HUMLIBST) probing for 96 cellular genes is commercially available from Compugen (http://www.labonweb.com). The HHV-8 60-65 nt oligomer set was custom designed by Compugen or in-house based on available genomic or cDNA sequence attainable from GenBank or additional published information, particularly for alternatively spliced messages (see Lu et al, publication data to be inserted). The human housekeeping and KSHV oligomer probe sets were distributed into 384-well polypropylene plates (Nalge Nunc Intl, 384 micro array) and dissolved in 3XSSC print buffer at a concentration of 200 ng/µl or 10µM. The 384-well plate contained either 3 wells with each housekeeping gene probe and 1 well with each KSHV probe, or duplicate wells for each housekeeping probe and duplicate wells for each KSHV probe. The microarray spot densities were 250 microns (center-to-center). Duplicate spots were printed per probe in two separate regions of each slide. The microarrays are printed on locally produced poly-L lysine coated slides and post-processed according to DNA microarrays: a molecular manual, 2003 edited by D. Bowtell and J. Sambrook, CSHL Press, Cold Spring Harbor, NY. Keywords = Human Herpesvirus-8 Keywords = HHV-8 Keywords = Kaposi's sarcoma-associated herpesvirus Keywords = KSHV Keywords = long oligomer microarray
Less...
More...