title: PMCC Human 10.5K
status: Public on Nov 27 2010
contact.city: Melbourne
contact.first: Holger
contact.last: Schlueter
contact.email: holger.schluter@petermac.org
contact.state: Victoria
contact.address: Locked Bag 1
contact.department: Research
contact.country: Australia
contact.web_link: http://www.petermac.org
contact.institute: Peter Mac Callum cancer Centre
contact.phone: 0061393299479
organism: Homo sapiens
manufacture_protocol: PRINTING: Arrays were printed on Telechem Superamine slides. The DNA concentration we print at is 200ng/ul. DNA concentrations in every well always differ. In the past have worked out the concentrations of random wells and assumed 8ug/100ul. PCR product is dried down and resuspended in 40ul to give 200ng/ul. One 384 well plate is made up of four 96 well plates.
  Cleaned up PCR product - 6ul is transferred to a 384 well plate, dried down then resuspended in 6ul of filtered 150mM NaPhosphate buffer pH 8 in a fridge overnight, ready for printing. After this, plates are resuspended in filtered Milli Q water. Plates are spun down in the centrifuge at a speed of 2800 rcf for 5 mins before resuspending in the fridge overnight, then placed on the shaker for 10 minutes then spun down again before printing. PCR foil seals were used to cover the 384 well plates during resuspension. After printing the arrays were dried down and covered with clear seal and stored in the -20C freezer. Pins take up ~0.25ul each time, so after a plate is printed, if dried down the plate was resuspended at 0.25ul less for the next print run.Scorecard aliqouts were resuspended in 5ul.
  PHOSPHATE BUFFER: For pH8: 95ml 0.5M Na (2) HPO (4) 5ml 0.5M NaH (2) PO (4). Water used for the array and for washing the pins is distilled water bought from Coles. Human 10.5K array. 4 x 6 = 24 pins = 16 pick ups per 384 well plate. Consists of 27 plates + scorecard = 28 plates