title: Cardiff NIA 15K set, print format 2
status: Public on Jul 22 2008
contact.city: Cardiff
contact.first: Fiona
contact.middle: Catherine
contact.last: Mansergh
contact.email: mansergf@tcd.ie
contact.state: Wales
contact.address: Museum Avenue
contact.department: Biosciences
contact.country: United Kingdom
contact.institute: Cardiff University
contact.phone: 0035318962484
organism: Mus musculus
description: Slides contain 16128 spots, printed in 12x4 mini grids, each with 16 rows and 21 columns. Spots representing NIA ESTs have the NIA clone number corresponding to their grid location. Some spaces on each grid are empty or blank.  Landmarks were used to align the grids, Scorecard/UT1/Rich spots are for use with a spike control. Spots actually marked as "empty" were printed with water/DMSO.
manufacture_protocol: Bacterial cells were grown in 96 well plates, pelleted at 2500rpm for 10 minutes, and resuspended in 50ul water. Cells were ruptured by incubation at 95 degrees celsius for 10 minutes, then spun for debris removal at 2500rpm for 10 minutes at 4 degrees. 10ul cell lysate was then added to 10ul 2xPCR mix (0.4ul 1omm dNTPs, 2ul each fwd and revs M13 primers (10pmol/ul, 2ul 10X Taq buffer, 2ul 25mM MgCl2, 0.4ul Taq polymerase (Promega), and 1.2ul sterile water) in a 96 well plate Amplification was carried out using an MWG thermocycler, 95oC, 1min then (95oC 1min, 58oC, 1min, 72oC, 30 secs). PCR products were then purified using Sephacryl S-300 and a 96 well filter plate (350ul/well). PCR products were assessed for successful amplification via 1.5% agarose gel electrophoresis. PCR products were then transferred to 384 well plates in known order then diluted to 50% original concentration with DMSO. PCR DNA samples were then used for printing on Corning CMT-GAPS slides.