title: Synechocystis sp. strain PCC 6803 12K full genome array version 4
status: Public on Oct 28 2010
contact.city: Dunedin
contact.first: Tina
contact.last: Summerfield
contact.email: tina.summerfield@otago.ac.nz
contact.address: 464 Great King Street
contact.department: Botany
contact.country: New Zealand
contact.web_link: http://www.botany.otago.ac.nz/people/summerfield.html
contact.institute: University of Otago
contact.phone: +64 3 4797578
organism: Synechocystis sp. PCC 6803
manufacture_protocol: Construction and Preparation of Synechocystis Spotted Microarrays: 3168 genes or putative ORFs of Synechocystis sp. PCC6803 were PCR amplified in two stages. Primary amplification was achieved using gene-specific up-stream and down-stream primers modified with adaptamer sequences and genomic DNA template. Secondary amplification was achieved using adaptamer primers and individual primary PCR product templates. The quality of the PCR products was determined by agarose gel electrophoresis. The PCR products were purified using Millipore Multi-screen plates and resuspended in Micro Spotting buffer (TeleChem; Sunnyvale, CA) to a concentration of 500ng/ul. Each probe was then printed (spotted) in triplicate onto TeleChem SuperAmine substrates at room temperature and 50% relative humidity using a GeneMachines Omnigrid 100 (Genomic Solutions; Ann Arbor, MI) robot fitted with 4x4 TeleChem SMP4 pins. After printing, each slide was rehydrated by holding the slide with the array side down over a container of steaming water for 1 s and snap-dried on a warm hot plate. Printed slides were baked at 80°C for 1 h and then subjected to a UV-cross-linking at a dose of 1500 mJ cm2. Slides with immobilized probes were then stored in slide holders and placed in a desiccator at room temperature.