title: Rice 2K cDNA microarray
status: Public on Mar 10 2009
contact.city: Wuhan
contact.first: Yuanyuan
contact.last: Wang
contact.email: cherry_wyy78@yahoo.com.cn
contact.address: College of Life Sciences, Wuhan University
contact.department: College of Life Sciences
contact.country: China
contact.institute: Wuhan University
contact.phone: 86 27 68752384
organism: Oryza sativa
manufacture_protocol: Subtractive cDNA library has been constructed from the BPH-infested B5 and MH63 plants using SSH technique (Yuan et al., 2005; Wang et al., 2005). To reduce the bias caused by SSH technology, equal numbers of clones were randomly selected from the each of the SSH cDNA libraries and a total of 1920 clones were used in the cDNA microarray analysis. The cDNA microarrays were custom-made and analyzed at TaKaRa (Dalian) Co., Ltd. To determine the detection sensitivity limit, the PCR-amplified fragment from lambda-A and Puc19 were used as external controls, and the PCR-amplified fragments from the mouse (Mus musculus) ß-actin gene and Human TFR gene were used as negative controls. Inserts of cDNA clones were PCR-amplified with T7 and M13R primers. The purified PCR products were dissolved in H2O at a concentration of 1.0 mg/ml and arrayed in paired dots from 384-well microtiter plates on poly-l-Lys-coated micro glass slides using a microarray stamping machine (Affymetrix 417 Arrayer, Affymetrix, Inc. Co, Santaclara, CA, USA). Such slides were post-processed according to the manufacturer’s protocols at TaKaRa Co., Ltd.