title: AGRF Human 8K cDNA Microarray (muscle study)
status: Public on Nov 01 2011
contact.city: Melbourne
contact.first: Christopher
contact.last: Bye
contact.state: Victoria
contact.address: University of Melbourne
contact.country: Australia
contact.institute: The Florey Institute of Neuroscience and Mental Health
organism: Homo sapiens
manufacture_protocol: H8K020304  Spotted  Corning GAPS-II Amino silane Slides  25x75mm Slides  Corning GAPS-II Amino silane Slides  16128 dots  Reporter Type:  PCR products of cDNA clones  Double stranded  Primer sequence used to amplify human 8K clone set plates 1-75  Reverse 5'gtgagcggataacaatttcacacaggaaacagc3'  Forward 5'ctgcaaggcgattaagttgggtaac 3'  Primer sequence used to amplify human 8K clone set plates 76-78  Reverse 5'cacaggaaacagctatgacca3'  Forward 5'acggccagtgccagcttat3'  cDNA Clone set from Research Genetics, Alabama, USA  Human 8K clone set obtained Nov 2001, Housed at AGRF, Melbourne Australia  Amplification Protocol: (Human 8K only)  0.2mM dNTP, 25 pmols each primer, 2.5 units HotStar Qiagen TAQ, 1X HotStar buffer, and 1ul straight clone culture in 100ul reactions.  95C for 15min; 7 cycles of 94C for 30s; 72C for 1min (-1C every cycle); 72C for 2min; 23 cycles of 94C for 30s; 65C for 1min; 72C for 2min; final elongation 72C for 5min; 4C 2min.  Millipore PCR plate purification, gel analysis, aliquot into 384 well plates by robot (A1,A2,B1,B2).  Resuspended in water for 4 days in fridge, then added an equal amount of DMSO.  Printing concentration was ~250 ng/ul in 50% DMSO, 6ul total.  Please see GAL file for sequence information and accession numbers for individual clones.  Post Processing: UV crosslinked 70mj; baked 2hrs at 80C; stored dessicated.  Feature Information:  ~150um diameter  Covalent and ionic attachment.  All other information, please see GAL file.  Control Elements on the array (see location on GAL file):  Amersham Lucinda Universal Scorecard version 2 spots. Spike mix available from the AGRF