title: Shewanella piezotolerans WP3 whole genome microarray
status: Public on Jan 30 2013
contact.city: Shanghai
contact.first: Huahua
contact.last: Jian
contact.email: jiandy@sjtu.edu.cn
contact.address: No.800 Dongchuan Road
contact.country: China
contact.institute: Shanghai Jiao Tong University
organism: Shewanella piezotolerans WP3
manufacture_protocol: 1. PCR primers for 4744 of 4945 predicted ORFs in the Shewanella piezotolerans WP3 genome (CP000472.1; excluding 200 CDSs shorter than 150bp) were designed by using Primer version 5.0 and then synthesized.
  2. ORF-specific fragments were amplified by using the following cycling conditions: denaturation for 30 s at 95℃, annealing for 1 min at 60℃ and extension for 1.5 min at 72℃, along with an initial 5 min denaturation at 95℃ and a final extension reaction at 72℃ for 10 min.
  3. All PCR products were purified by using ethanol precipitation. The quality of the amplified products was checked by 1.5% agarose gel electrophoresis and ethidium bromide staining.
  4. PCR products and microarray reagents were arrayed from a 384-well microtiter plate. Probes were printed in triplicate onto Telechem Superamine slides (Telechem, Sunnyvale, USA). The printed slides were dried and subjected to UV cross-linking.