title: Shanghai Biochip Staph epidermidis 48 oligo array
status: Public on Mar 08 2010
contact.city: Shanghai
contact.first: Tao
contact.last: Zhu
contact.email: 071101036@fudan.edu.cn
contact.address: 138, Dongan Road
contact.country: China
contact.institute: Fudan university
organism: Staphylococcus epidermidis
manufacture_protocol: 1. 50-mer oligos designed according to the S. epidermidis RP62A genome sequence were synthsized.
  2. Poly-L-lysine-coated glass slides of 75 × 25 mm were prepared as described (http://cmgm.stanford.edu/pbrown/MGuide). Slides with aminosilane surface (CMT-GAPS™) were purchased from Corning (Corning, USA). The DNA spotting solution was adjusted to either 45 mM sodium citrate pH 7.0, 450 mM NaCl (3× SSC) or the same composition supplemented with 1.5 M betaine (N,N,N-trimethylglycine; Sigma, Germany).
  3. DNA spotting was done with an SDDC-2 DNA Micro-Arrayer from Engineering Services Inc. (Toronto, Canada). A single SMP3 pin (TeleChem International Inc., Sunnyvale, USA) was used to avoid differences between pins. The DNA samples were printed in quadruplicate at a 200 µm centre-to-centre spacing. Slides were left at room temperature overnight and then heat-treated on a metal block at 80°C for 5 s.
  4. The DNA was crosslinked to the support by UV irradiation with a total energy of 60 mJ in a Hoefer UV-crosslinker (Amersham Pharmacia Biotech, Freiburg, Germany).
  5. For the blocking process, 1 g succinic anhydride (Fluka, Deisendorf, Germany) was freshly dissolved in 200 ml anhydrous 1,2-dichloroethane (DCE; Fluka). To this solution, 2.5 ml of N-methylimidazol (Fluka) was added and immediately poured into the slide chamber. Incubation was for 1 h, placed on an orbital shaker for slight agitation.
  6. Subsequently, the slides were briefly washed in 200 ml of fresh DCE and incubated in boiling water for 2 min for DNA denaturation. After a brief rinse in 95% ethanol, they were left to dry at room temperature.